Pharmaceutical compounds

ABSTRACT

Fused pyrimidines of formula (I): 
     
       
         
         
             
             
         
       
     
     wherein R 1 -R 3 , A and n have any of the values described in the specification; and pharmaceutically acceptable salts thereof, have activity as inhibitors of PI3K and may thus be used to treat diseases and disorders arising from abnormal cell growth, function or behaviour associated with PI3 kinase such as cancer, immune disorders, cardiovascular disease, viral infection, inflammation, metabolism/endocrine disorders and neurological disorders. Processes for synthesizing the compounds are also described.

PRIORITY OF THE INVENTION

This application is a Continuation of PCT/GB2005/004129, filed Oct. 25,2005, which claims priority from UK Application No. 0423653.5, filedOct. 25, 2004, which applications are incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to pyrimidine derivatives and their use asinhibitors of phosphatidylinositol 3-kinase (PI3K).

BACKGROUND TO THE INVENTION

Phosphatidylinositol (hereinafter abbreviated as “PI”) is one of anumber of phospholipids found in cell membranes. In recent years it hasbecome clear that PI plays an important role in intracellular signaltransduction. In the late 1980s, a PI3 kinase (PI3K) was found to be anenzyme which phosphorylates the 3-position of the inositol ring ofphosphatidylinositol (D. Whitman et al, 1988, Nature, 332, 664).

PI3K was originally considered to be a single enzyme, but it has nowbeen clarified that a plurality of subtypes are present in PI3K. Eachsubtype has its own mechanism for regulating activity. Three majorclasses of PI3Ks have been identified on the basis of their in vitrosubstrate specificity (B. Vanhaesebroeck, 1997, Trend in Biol. Sci, 22,267). Substrates for class I PI3Ks are PI, PI 4-phosphate (PI4P) and PI4,5-biphosphate (PI (4,5)P2). Class I PI3Ks are further divided into twogroups, class Ia and class Ib, in terms of their activation mechanism.Class Ia PI3Ks include PI3K p110α, p110β and p110δ subtypes, whichtransmit signals from tyrosine kinase-coupled receptors. Class Ib PI3Kincludes a p110γ subtype activated by a G protein-coupled receptor. PIand PI(4)P are known as substrates for class II PI3Ks. Class II PI3Ksinclude PI3K C2α, C2β and C2γ subtypes, which are characterized bycontaining C2 domains at the C terminus. The substrate for class IIIPI3Ks is PI only.

In the PI3K subtypes, the class Ia subtype has been most extensivelyinvestigated to date. The three subtypes of class Ia are heterodimers ofa catalytic 10 kDa subunit and regulatory subunits of 85 kDa or 55 kDa.The regulatory subunits contain SH2 domains and bind to tyrosineresidues phosphorylated by growth factor receptors with a tyrosinekinase activity or oncogene products, thereby inducing the PI3K activityof the p110 catalytic subunit which phosphorylates its lipid substrate.Thus, the class Ia subtypes are considered to be associated with cellproliferation and carcinogenesis.

WO 01/083456 describes a series of condensed heteroaryl derivativeswhich have activity as inhibitors of PI3K and which suppress cancer cellgrowth.

SUMMARY OF THE INVENTION

It has now been found that a novel class of fused pyrimidine compoundsare effective inhibitors of PI3K with drug-like physicochemical andpharmacokinetic properties. The compounds exhibit selectivity for classIa PI3Ks over class Ib, in particular for the P110α subtype.

Accordingly, the present invention provides a compound which is a fusedpyrimidine of formula (I):

whereinA represents a thiophene or furan ring;n is 1 or 2;R¹ is a group of formula:

whereinm is 0 or 1;R³⁰ is H or C₁-C₆ alkyl;R⁴ and R⁵ form, together with the N atom to which they are attached, a5- or 6-membered saturated N-containing heterocyclic group whichincludes 0 or 1 additional heteroatoms selected from N, S and O, whichmay be fused to a benzene ring and which is unsubstituted orsubstituted; or one of R⁴ and R⁵ is alkyl and the other is a 5- or6-membered saturated N-containing heterocyclic group as defined above oran alkyl group which is substituted by a 5- or 6-membered saturatedN-containing heterocyclic group as defined above;R² is selected from:

-   -   (a)

-   -   wherein R⁶ and R⁷ form, together with the nitrogen atom to which        they are attached, a morpholine, thiomorpholine, piperidine,        piperazine, oxazepane or thiazepane group which is unsubstituted        or substituted; and    -   (b)

-   -   wherein Y is a C₂-C₄ alkylene chain which contains, between        constituent carbon atoms of the chain and/or at one or both ends        of the chain, 1 or 2 heteroatoms selected from O, N and S, and        which is unsubstituted or substituted;        and R³ is an indazole group which is unsubstituted or        substituted;        or a pharmaceutically acceptable salt thereof.

DETAILED DESCRIPTION OF THE INVENTION

The thiophene or furan ring A in formula (I) adopts either of the twoavailable regiochemical orientations. Formula (I) thus covers thethieno[3,2-d]pyrimidines and furano[3,2-d]pyrimidines of the followingformula (Ia) as well as the thieno[2,3-d]pyrimidines andfurano[2,3-d]pyrimidines of the following formula (Ib):

wherein each of R¹ to R³ and n is as defined above and X is S or O.

In formula (I), the group or groups R¹, which are the same or differentin a given compound when n is 2, may be bonded to either or both of thetwo available ring positions on the thiophene or furan ring A. Referringto structures (Ia) and (Ib) above, therefore, when n is 1 the furan orthiophene ring is mono-substituted by R¹ at the 2-position or the3-position. When n is 2, the thiophene or furan ring is di-substitutedby R¹ at positions 2 and 3.

As specified herein, an alkyl group is a straight or branched chainsaturated hydrocarbon radical which is unsubstituted or substituted.Typically it is C₁-C₂₀ alkyl, for instance C₁-C₁₀ alkyl, such as C₁-C₆alkyl or C₁-C₄ alkyl, for example methyl, ethyl, i-propyl, n-propyl,t-butyl, s-butyl or n-butyl. It may also be pentyl, hexyl, heptyl, octyland the various branched chain isomers thereof.

When an alkyl group is substituted it typically bears one or moresubstituents R²⁰ selected from halogen, alkoxy, carbocyclyl, a 5- or6-membered saturated N-containing heterocyclic group as defined above,OH, SR, CN, nitro, NR₂, —COOR, —C(O)R, —CH₂OR, S(O)_(m)R, —NRC(O)R,—S(O)_(m)NR₂, —OC(O)R, —OC(O)NR₂, —NRS(O)_(m)R, —NRC(O)NR₂ and —CONR₂,wherein each R is H, unsubstituted alkyl or C₃-C₁₀ cycloalkyl and m is 1or 2.

Typically R²⁰ is selected from halogen, alkoxy, carbocyclyl, a 5- or6-membered saturated N-containing heterocyclic group as defined above,OH, CN, NR₂, —COOR and —CONR₂, wherein each R is H or unsubstitutedalkyl as defined above.

Substituted alkyl may be, for instance, a haloalkyl group or a group-alk-N(R⁴)(R⁵) wherein alk is an alkylene chain and R⁴ and R⁵ form,together with the N atom to which they are attached, a 5- or 6-memberedsaturated N-containing heterocyclic group which includes 0 or 1additional heteroatoms selected from N, S and O, which may be fused to abenzene ring and which is unsubstituted or substituted. More typicallyit is a haloalkyl group or a group -alk-N(R⁴)(R⁵) wherein alk is analkylene chain and R⁴ and R⁵ form, together with the N atom to whichthey are attached, a 5- or 6-membered saturated N-containingheterocyclic group as defined above.

An alkylene group is unsubstituted or substituted, straight or branchedchain saturated divalent hydrocarbon group. Typically it is C₁-C₈alkylene, for instance C₁-C₆ alkylene. Preferably it is C₁-C₄ alkylene,for example C₂-C₄ alkylene, such as methylene, ethylene, i-propylene,n-propylene, t-butylene, s-butylene or n-butylene. It may also bepentylene, hexylene, heptylene, octylene and the various branched chainisomers thereof. When the alkylene group is substituted it is typicallysubstituted by a group R²⁰ as defined above or by alkyl which isunsubstituted or substituted by a group R²⁰ as defined above.

An alkenyl group is an unsubstituted or substituted, straight orbranched chain hydrocarbon radical having one or more double bonds.Typically it is C₂-C₈ alkenyl, for instance C₂-C₆ alkenyl, such asallyl, butenyl, butadienyl, pentenyl or hexenyl. When the alkenyl groupis substituted it is typically substituted by a group R²⁰ as definedabove or by alkyl which is unsubstituted or substituted by a group R²⁰as defined above.

An alkynyl group is an unsubstituted or substituted, straight orbranched chain hydrocarbon radical having one or more triple bonds.Typically it is C₂-C₈ alkynyl, for instance C₂-C₆ alkynyl, such asethynyl, propynyl or butynyl. When the alkynyl group is substituted itis typically substituted by a group R²⁰ as defined above or by alkylwhich is unsubstituted or substituted by a group R²⁰ as defined above.

A haloalkyl group is an alkyl group as defined above, substituted by oneor more halogen atoms. It can be a perhaloalkyl group, for instancetrifluoromethyl or perfluorohexyl.

A halogen is chlorine, fluorine, bromine or iodine. It is typicallybromine or iodine.

An alkoxy group is straight or branched chain. It is typically C₁-C₆alkoxy, for instance C₁-C₄ alkoxy, such as methoxy, ethoxy, i-propoxy,n-propoxy, t-butoxy, n-butoxy or s-butoxy. It is unsubstituted orsubstituted, for instance by a group R²⁰ as defined above or by alkylwhich is unsubstituted or substituted by a group R²⁰ as defined above.Typically it is substituted by carbocyclyl, morpholino, OH, CN, NR₂,—COOR or —CONR₂, wherein each R is H or unsubstituted alkyl as definedabove.

A carbocyclyl group is a non-aromatic saturated or unsaturatedmonocyclic hydrocarbon ring, typically having from 3 to 10 carbon atoms.It may be a C₃-C₈ cycloalkyl group, or C₅-C₁₀ cycloalkyl group, forinstance cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptylor cyclooctyl. Alternatively it may be a cycloalkenyl group, typicallyC₄-C₈ cycloalkenyl, for instance cylcopentenyl, cyclohexenyl,cyclohexadienyl, cycloheptenyl, cyclohepadienyl, cyclooctenyl orcyclooctadienyl. A carbocyclyl group may be unsubstituted orsubstituted, for instance by a group R²⁰ as defined above or by alkylwhich is unsubstituted or substituted by a group R²⁰ as defined above.Typically it is substituted by alkoxy, morpholino, OH, CN, NR₂, —COOR or—CONR₂, wherein each R is H or unsubstituted alkyl as defined above.

A 5- or 6-membered saturated N-containing heterocyclic group whichincludes 0 or 1 additional heteroatoms selected from N, S and O, whichmay be fused to a benzene ring and which is unsubstituted or substitutedis typically selected from morpholine, piperidine, piperazine,pyrrolidine, thiomorpholine, quinoline, isoquinoline, diazepane,oxazepane and thiazepane.

When a 5- or 6-membered saturated N-containing heterocyclic group asdefined above is substituted it is typically substituted by one or moresubstituents, for instance 1, 2 or 3 substituents, typically by 1 or 2substituents. Typically the substituents are selected from alkyl whichis unsubstituted or substituted, alkoxy which is unsubstituted orsubstituted, —NR₂, —N(R′″)-alk-OR, -alk-OR, —O-alk-OR, -alk-C(O)NR₂,—C(O)NR₂, -alk-Het, —N(R)-Het, —O-Het, —N(R)—C(O)-alk-OR,—C(O)—N(R)-alk-OR, -alk-S(O)₂R, —N(R)-alk-OR, -alk-NR′R″,—N(R′″)—S(O)₂R, S(O)₂R′″, -alk-N(R)-alk-OR, —S(O)₂-alk-OR, a second 5-or 6-membered saturated N-containing heterocyclic group as definedabove, a 5- or 6-membered N-containing heteroaryl group which isunsubstituted or substituted and which may be fused to a benzene ring,—COOR, —CONR₂, oxo (═O), —SO₂NR₂, —SO₂-alk-NR₂ and —CO-alk-OR, wherein:alk is an alkylene chain as defined above; Het is a 5- or 6-memberedN-containing heteroaryl group as defined herein which is unsubstitutedor substituted; R is H or alkyl, or when two groups R are bonded to Nthey may form, together with the N atom, a saturated 5- or 6-memberedN-containing heterocyclic group as defined herein which is unsubstitutedor substituted; each of R′ and R″ is independently H, alkyl or alkoxy;and R′″ is alkyl which is unsubstituted or substituted, for instance byCF₃, NR₂, OR, a 5- or 6-membered saturated N-containing heterocyclicgroup as defined herein or a 5- or 6-membered N-containingheteroaryl-group as defined herein, the said heterocyclic and heteroarylgroups being unsubstituted or substituted.

A 5-, 6- or 7-membered saturated heterocyclic group which contains 1 or2 heteroatoms selected from N, S and O and which is unsubstituted orsubstituted is typically selected from tetrahydropyran,tetrahydrothiopyran, tetrahydrofuran and tetrahydrothiofuran.

When a 5-, 6- or 7-membered saturated heterocyclic group which contains1 or 2 heteroatoms selected from N, S and O is substituted it may besubstituted by a group R²⁰ as defined above. Typically it is substitutedby one or more substituents selected from alkyl as defined above whichis unsubstituted or substituted, for instance by R²⁰ as defined above orby alkyl which is unsubstituted or substituted by a group R²⁰ as definedabove, haloalkyl as defined above, alkoxy as defined above which isunsubstituted or substituted, halogen, hydroxy, CN, nitro, amino, oxo(═O), and —NR′R″ wherein each of R′ and R″ is independently H or alkyl.

A heteroaryl group is a heteroaryl group which contains 1, 2, 3 or 4ring nitrogen atoms and 0, 1 or 2 additional heteroatoms selected fromO, N and S, which group is monocyclic or bicyclic and which isunsubstituted or substituted. It is typically a 5- to 12-membered ring.Examples of a heteroaryl group include pyrrole, pyrazole, triazole,tetrazole, indazole, thiazole, isothiazole, oxazole, isooxazole, indole,isoindole, 1,3-dihydro-indol-2-one, pyridine-2-one, pyridine,pyridin-3-ol, imidazole, 1,3-dihydro-benzimidazolone, benzimidazole,benzothiazole, benzothiadiazole, quinoline, isoquinoline, quinoxaline,pyrazolopyridine, aminopyrazolinone, imidazopyridine, pyrimidine,pyridazine, pyrazine and isatin groups. Preferred examples includeindazole, indole, pyrazole and tetrazole groups. These groups may beunsubstituted or substituted, for instance by a group R²⁰ as specifiedabove or by alkyl which is unsubstituted or substituted by a group R²⁰as defined above.

A 5- or 6-membered N containing heteroaryl group which may be fused to abenzene ring is typically selected from pyrrole, pyrazole, triazole,tetrazole, indazole, thiazole, isothiazole, oxazole, isooxazole, indole,isoindole, 1,3-dihydro-indol-2-one, pyridine-2-one, pyridine,pyridin-3-ol, imidazole, 1,3-dihydro-benzimidazolone, benzimidazole,benzothiazole, benzothiadiazole, quinoline, isoquinoline, quinoxaline,pyrazolopyridine, aminopyrazolinone, imidazopyridine, pyrimidine,pyridazine and pyrazine. When such a heteroaryl group is substituted itmay be substituted by a group R²⁰ as defined above or by alkyl which isunsubstituted or substituted by a group R²⁰ as defined above.

In R¹, m is 0 or 1, typically 1. R³⁰ is typically H. R⁴ and R⁵ typicallyform, together with the N atom to which they are attached, a saturatedN-containing heterocyclic group selected from morpholine,thiomorpholine, piperidine, piperazine, pyrrolidine, quinoline,isoquinoline, diazepane, oxazepane and thiazepane. The heterocylic groupformed by R⁴ and R⁵ is unsubstituted or substituted, for instance by oneor more substituents selected from alkyl which is unsubstituted orsubstituted, alkoxy which is unsubstituted or substituted,—N(R′″)-alk-OR, -alk-OR, —O-alk-OR, -alk-C(O)NR₂, —C(O)NR₂, -alk-Het,—N(R)-Het, —O-Het, —N(R)—C(O)-alk-OR, —NR—S(O)₂R, —N(R)-alk-S(O)₂R,—N(R)-alk-OR, -alk-NR′R″, —N(R′″)—S(O)₂R, S(O)₂R′″, -alk-N(R)-alk-OR,—S(O)₂-alk-OR and a 5- or 6-membered N-containing heteroaryl group whichis unsubstituted or substituted and which may be fused to a benzenering, —COOR, —CONR₂, oxo (═O), —SO₂NR₂, —SO₂-alk-NR₂ and —CO-alk-OR,wherein: alk is an alkylene chain as defined above; Het is a 5- or6-membered N-containing heteroaryl group as defined herein which isunsubstituted or substituted; R is H or alkyl, or when two groups R arebonded to N they may form, together with the N atom, a saturated 5- or6-membered N-containing heterocyclic group as defined herein which isunsubstituted or substituted; each of R′ and R″ is independently H,alkyl or alkoxy; and R′″ is alkyl which is unsubstituted or substituted,for instance by CF₃, NR₂, OR, a 5- or 6-membered saturated N-containingheterocyclic group as defined herein or a 5- or 6-membered N-containingheteroaryl group as defined herein, the said heterocyclic and heteroarylgroups being unsubstituted or substituted.

In this definition of R¹ Het is typically selected from pyridine (forinstance pyridin-1-yl, pyridin-2-yl or pyridin-3-yl), pyrimidine,imidazole, furan, oxazole, isoxazole and thiazole, each of which isunsubstituted or substituted. The moiety “alk” is typically astraight-chain C₁-C₄ alkylene group, more typically C₁-C₃ alkylene, suchas —CH₂—, —CH₂CH₂, or —CH₂CH₂CH₂—.

In definition (a) of R² in formula (I), the ring formed by R⁶ and R⁷ istypically morpholine which is unsubstituted or substituted, for instanceby a group R²⁰ as specified above. It may alternatively be a groupselected from tetrahydropyran, tetrahydrothiopyran, tetrahydrofuran andtetrahydrothiofuran, each of which is unsubstituted or substituted, forinstance, for instance by a group R²⁰ as specified above. When the ringformed by R⁶ and R⁷ is substituted it may be substituted on either aring heteroatom or a ring carbon atom, for instance by a group R²⁰ asdefined above or by alkyl which is unsubstituted or substituted by agroup R²⁰ as defined above.

In definition (b) of R² in formula (I), the alkylene chain representedby Y forms, together with the carbon atoms to which it is attached, asaturated 5-, 6- or 7-membered heterocyclic ring which contains 1 or 2heteroatoms selected from O, N and S and which is unsubstituted orsubstituted. Examples of the heterocyclic ring include tetrahydropyran,tetrahydrofuran, tetrahydrothiopyran, tetrahydrothiofuran andmorpholine. When the heterocyclic ring is substituted it is typicallysubstituted by one or more substituents, for instance 1, 2 or 3substituents, selected from halogen, alkyl, haloalkyl (for instancetrifluoromethyl), alkoxy, OH, CN, NR₂, oxo (═O), —COOR and —CONR₂,wherein each R is H or unsubstituted alkyl as defined above.

The indazole group in the definition of R³ is unsubstituted orsubstituted. If it is substituted it may be substituted by one or moresubstituents selected from a group Z, wherein Z is selected from OR,CH₂OR, CO₂R, CF₂OH, CH(CF₃)OH, C(CF₃)₂OH, —(CH₂)_(q)OR and —(CH₂)_(q)NR₂wherein each R is independently H or alkyl and q is 0, 1 or 2; one ormore substituents selected from halo, alkyl, alkenyl, alkynyl, CN, NO₂,OR, SR, NR₂, C(O)R, SOR, SO₂R, SO₂NR₂, NC(O)R and CO₂R, wherein each Ris independently H or alkyl; and an oxo group (═O). Typically, ifsubstituted, the indazole group is substituted by OH, NH₂ or an oxogroup. In one embodiment the indazole group is unsubstituted.

The indazole group R³ is an isostere of a 3-hydroxyphenyl or4-hydroxyphenyl group. An isostere as used herein is a functional groupwhich possesses binding properties which are the same as, or similar to,the 3-hydroxyphenyl or 4-hydroxyphenyl group in the context of thestructure of formula (I).

In one embodiment the fused pyrimidine is of formula (Ic):

whereinR² and R³ are as defined above;n is 1; andR¹ is a group of formula:

whereinm is 0 or 1;R⁴ and R⁵ form, together with the N atom to which they are attached, a5- or 6-membered saturated N-containing heterocyclic group whichincludes 0 or 1 additional heteroatoms selected from N, S and O, andwhich is unsubstituted or substituted by one or more substituentsselected from alkyl which is unsubstituted or substituted, alkoxy whichis unsubstituted or substituted, —NR₂, —N(R′″)-alk-OR, -alk-OR,—O-alk-OR, -alk-C(O)NR₂, —C(O)NR₂, -alk-Het, —N(R)-Het, —O-Het,—N(R)—C(O)-alk-OR, —C(O)—NR-alk-OR, -alk-S(O)₂R, —N(R)-alk-OR,-alk-NR′R″, —N(R′″)—S(O)₂R, S(O)₂R′″, -alk-N(R)-alk-OR, —S(O)₂-alk-OR,and a 5- or 6-membered N-containing heteroaryl group which isunsubstituted or substituted and which may be fused to a benzene ring,wherein alk is an alkylene chain as defined above, Het is a 5- or6-membered N-containing heteroaryl group as defined herein which isunsubstituted or substituted, R is H or alkyl or when two groups R arebonded to N they may form, together with the N atom, a saturated 5- or6-membered N-containing heterocyclic group as defined herein which isunsubstituted or substituted; each of R′ and R″ is independently H,alkyl or alkoxy, and R′″ is alkyl which is unsubstituted or substituted,or one of R⁴ and R⁵ is alkyl and the other is a 5- or 6-memberedsaturated N-containing heterocyclic group as defined above, which isunsubstituted or substituted as defined above, or an alkyl group whichis substituted by a 5- or 6-membered saturated N-containing heterocyclicgroup as defined above;or a pharmaceutically acceptable salt thereof.

In formula (Ic) the moiety “alk” is typically a straight-chain C₁-C₄alkylene group, more typically C₁-C₃ alkylene, such as —CH₂—, —CH₂CH₂,or —CH₂CH₂CH₂—. The heterocyclic group formed by R⁴ and R⁵ is typicallyselected from morpholine, piperidine and piperazine, each of which isunsubstituted or substituted as defined above. R² is typicallymorpholine. R³ is typically an indazole group which is unsubstituted.

Specific examples of compounds of the invention include:

-   2-(1H-Indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-sulfonic    acid dimethylamide;-   {4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-morpholin-4-yl-methanone;-   4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-carboxylic    acid (2-methoxy-ethyl)-methyl-amide;-   {4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-N,N-dimethyl-acetamide;-   4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-carboxylic    acid dimethylamide;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(3-morpholin-4-yl-propane-1-sulfonyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-methyl-amine;-   (3-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-sulfonyl}-propyl)-dimethyl-amine;-   2-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-2-methyl-propan-1-ol;-   1′-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-[1,4′]bipiperidinyl;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-morpholin-4-yl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyrimidin-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   1-(2-Hydroxy-ethyl)-4-[2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-2-one;-   6-(4-Cyclopropylmethyl-piperazin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-thiazol-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   2-(6-Fluoro-1H-indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-2-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-thiazol-2-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-[4-(5-methyl-furan-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylic    acid amide;-   2-(1H-Indazol-4-yl)-6-[4-(2-methoxy-1,1-dimethyl-ethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-[(3R,5S)-4-(2-methoxy-ethyl)-3,5-dimethyl-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylic    acid (2-methoxy-ethyl)-methyl-amide;-   1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylic    acid dimethylamide;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-3-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylic    acid methylamide;-   2-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-N-methyl-isobutyramide;-   )    2-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-2-methyl-1-pyrrolidin-1-yl-propan-1-one;-   2-(1H-Indazol-4-yl)-6-[4-(1-methyl-1H-imidazol-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-[4-(5-methyl-isoxazol-3-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   1-{4-[2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2    d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-2-methyl-propan-2-ol;-   Cyclopropylmethyl-{1-[2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-amine;-   6-[4-(1-Ethyl-1-methoxymethyl-propyl)-piperazin-1-ylmethyl]-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-[4-(1-methoxymethyl-cyclopropyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-(2,2,2-trifluoro-ethyl)-amine;-   2-(1H-Indazol-4-yl)-6-[4-(2-methoxy-ethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-methanol;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-4-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-[4-(6-methyl-pyridin-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-[4-(4-methyl-thiazol-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-pyridin-2-yl-amine;-   N-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-2-methoxy-N-methyl-acetamide;-   N-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-N-methyl-methanesulfonamide;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(3-methoxy-propyl)-methyl-amine;-   6-((3S,5R)-3,5-Dimethyl-4-pyridin-2-ylmethyl-piperazin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-(4-methoxymethyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-thiazol-2-ylmethyl-amine;-   1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-4-pyridin-2-ylmethyl-piperidin-4-ol;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-isopropyl-(2-methoxy-ethyl)-amine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(pyridin-2-yloxy)-piperidin-1-ylmethyl]-thieno[3,2-d]pyrimidine;-   N-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-N-(2-methoxy-ethyl)-methanesulfonamide;-   2-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-propan-2-ol;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(1-oxy-pyridin-3-ylmethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-morpholin-4-ylmethyl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-ylmethyl}-(2-methoxy-ethyl)-methyl-amine;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-ylmethyl}-dimethyl-amine;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-3-yl}-(2-methoxy-ethyl)-methyl-amine;-   1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-3-carboxylic    acid methylamide;-   2-(1H-Indazol-4-yl)-6-(3-methoxymethyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-2-ylmethyl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-[4-(2-methoxy-ethoxy)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   6-((3R,5S)-3,5-Dimethyl-4-thiazol-2-ylmethyl-piperazin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(1-oxy-pyridin-2-ylmethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-[4-(2-methoxy-ethyl)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-(4-methanesulfonyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(3-methanesulfonyl-propyl)-methyl-amine;-   2-(1H-Indazol-4-yl)-6-[4-(3-methoxy-propane-1-sulfonyl)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine.-   (R)-1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-3-carboxylic    acid methylamide;-   (S)-1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-3-carboxylic    acid methylamide;-   6-(4-Imidazol-1-ylmethyl-piperidin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-morpholin-4-ylmethyl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-6-(3-methyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   {1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-3-yl}-methanol;-   2-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-ethanol;-   1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-4-thiazol-2-yl-piperidin-4-ol;-   2-(1-Methyl-1H-indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(2-Methyl-2H-indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-thiazol-4-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   1-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-3-phenoxy-propan-2-ol;-   6-[4-(1H-Imidazol-2-ylmethyl)-piperazin-1-ylmethyl]-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   6-[4-(3H-Imidazol-4-ylmethyl)-piperazin-1-ylmethyl]-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;-   2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-((2S,6R)-2,4,6-trimethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;-   {4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-1-methanesulfonyl-piperazin-2-yl}-methanol;    and-   2-(1H-Indazol-4-yl)-6-(4-methanesulfonyl-3-methoxymethyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;    and the pharmaceutically acceptable salts thereof.

The compounds of formula (I) may exist in the form of geometricalisomers or tautomers depending on the kinds of substituent groups, andthese isomers in separated forms or mixtures thereof may be used in thepresent invention. Where the compounds have asymmetric carbon atoms,optical isomer forms may exist based on such carbon atoms. All of themixtures and the isolated forms of these optical isomers may be used inthe present invention.

A suitable synthetic strategy for producing compounds of formula (I) inwhich m is 1 employs the precursor carboxaldehyde of formula (II):

wherein A and R² are as defined above. Starting from this precursor thesynthesis comprises performing, in either order, a palladium-mediated(Suzuki-type) cross-coupling reaction and a reductive amination. Thepresent invention therefore further provides a process for producing acompound of formula (I) as defined above in which m is 1, which processcomprises:

(a) treating a compound of formula (II):

wherein A and R² are as defined above, with a boronic acid or esterthereof of formula R³B(OR¹⁵)₂, in which R³ is as defined above and eachR¹⁵ is H or C₁-C₆ alkyl or the two groups OR¹⁵ form, together with theboron atom to which they are attached, a pinacolato boronate estergroup, in the presence of a Pd catalyst; and treating the resultingcompound of formula (III):

wherein A, R² and R³ are as defined above, with an amine of formulaNHR⁴R⁵ in which R⁴ and R⁵ are as defined above, in the presence of asuitable reducing agent; or

(b) treating a compound of formula (II) as defined above with an amineof formula NHR⁴R⁵ wherein R⁴ and R⁵ are as defined above, in thepresence of a suitable reducing agent; and treating the resultingcompound of formula (IV):

wherein A, R², R⁴ and R⁵ are as defined above, with a boronic acid orester thereof of formula R³B(OR¹⁵)₂, in which R³ is as defined above andeach R¹⁵ is H or C₁-C₆ alkyl or the two groups OR¹⁵ form, together withthe boron atom to which they are attached, a pinacolato boronate estergroup, in the presence of a Pd catalyst.

Both the amination step and the Pd-mediated cross-coupling step takeplace under conventional conditions. The palladium catalyst may be anythat is typically used for Suzuki-type cross-couplings, such asPdCl₂(PPh₃)₂. The reducing agent is typically a borohydride, forinstance NaBH(OAc)₃, NaBH₄ or NaCNBH₄, in particular NaBH(OAc)₃.

The pinacolato boronate ester may be, for instance, prepared asdescribed in either of Reference Examples 5 and 6 which follow.

A compound of formula (II) as defined above wherein R² is —NR⁶R⁷ may beprepared by a process which comprises treating a compound of formula(IX):

wherein A, R⁶ and R⁷ are as defined above, with a lithiating agentfollowed by N,N′-dimethylformamide (DMF). The reaction is typicallyconducted by adding a solution of the lithiating agent in a non-polarorganic solvent, for instance a hydrocarbon solvent such as hexane, to asuspension of the compound of formula (IX) in an organic solvent such astetrahydrofuran (THF). If THF is used the addition takes place at a lowtemperature, of about −78° C. The lithiating agent is typically analkyllithium, for instance n-butyllithium.

A compound of formula (IX) as defined above may be produced by a processwhich comprises treating a compound of formula (X):

with an amine of formula NHR⁶R⁷, wherein R⁶ and R⁷ are as defined above,in an organic solvent. The solvent is typically an alcohol, such asmethanol. The reaction is generally conducted at room temperature.

A compound of formula (X) may be prepared by the process described inReference Example 1 for the preparation of2,4-dichloro-thieno[3,2-d]pyrimidine, or by analogy with such a process.

A compound of formula (II) as defined above wherein R² is of formula

may be prepared by a process which comprises submitting a compound offormula

wherein A and R³ are as defined above, to palladium-mediatedcross-coupling with a compound of formula (XII):

wherein L is H or a group selected from halo, —OSO₂CF₃, —B(OR)₂, —Sn(R)₃and —Si(R)₃ wherein R is H or alkyl as defined above, followed byreduction, to yield a compound of the following formula (XIII):

wherein A, R³ and Y are as defined above.

The compound of formula (XIII) may be converted to the correspondingcarboxaldehyde by treatment with a lithiating agent followed byN,N′-dimethylformamide (DMF), for instance under the conditionsdescribed above for the conversion of a compound of formula (IX) to acompound of formula (II). The lithiating agent is typically as definedabove. The resulting carboxaldehyde may then be converted into a desiredfinal compound of formula (I) as defined above, in which m is 1, bytreatment with an amine of formula NHR⁴R⁵ in which R⁴ and R⁵ are asdefined above, in the presence of a suitable reducing agent, forinstance a borohydride as specified above, in particular NaBH(OAc)₃.

A compound of formula (I) as defined above in which m is 0 may beprepared by a Buchwald-type palladium-mediated nitrogen insertionreaction. Such a process may comprise treating a compound of formula(XIV):

wherein A, R² and R³ are as defined above and W is a halo group selectedfrom Br and I, with an amine of formula NHR⁴R⁵ in which R⁴ and R⁵ are asdefined above, in the presence of a palladium catalyst.

A compound of formula (XIV) may be produced by treating a compound offormula (XV):

wherein A, R² and R³ are as defined above, with a lithiating agent and ahalogen selected from bromine and iodine. The lithiating agent istypically an alkyllithium, for instance butyllithium. The halogen istypically iodine, which gives rise to a compound of formula (XIV) inwhich W is I.

A compound of formula (I) as defined above in which m is 0 may also beprepared by an SNAr displacement reaction, for instance under theconditions described by D. Prim and G. Kirsch in Tetrahedron 1999, 55(21), 6511-6526. Such a process comprises treating a compound of formula(XIV) as defined above in which W is Br with an amine of formula NHR⁴R⁵in which R⁴ and R⁵ are as defined above in H₂O under reflux for 12 h.

A compound of formula (I) as defined above in which m is 0 mayalternatively be prepared by treating a compound of formula (XIV) asdefined above in which W is I with an amine of formula NHR⁴R⁵ in whichR⁴ and R⁵ are as defined above in 1,4-dioxane in the presence of CuI/Enand K₃PO₄. The reaction is conducted at about 110° C. for 24 h. Thisprocedure is described by Kang S-K et al in Synlett, (3), 427-430, 2002.

A fused pyrimidine of formula (I) may be converted into apharmaceutically acceptable salt, and a salts may be converted into thefree compound, by conventional methods. Examples of pharmaceuticallyacceptable salts include acid addition salts with inorganic acids suchas hydrochloric acid, hydrobromic acid, hydroiodic acid, sulphuric acid,nitric acid and phosphoric acid; and organic acids such as formic acid,acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonicacid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid,tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid,aspartic acid and glutamic acid. In the case of compounds of theinvention bearing a free carboxy substituent, the salts include thesalts of alkali and alkaline earth metals and ammonium, for instance thesalts of sodium, potassium, magnesium, calcium and ammonium. The latterare prepared by treating the free fused pyrimidine of formula (I), or anacid addition salt thereof, with the corresponding metal base orammonia. The compounds of formula (I) and their salts may exist ashydrates or solvates.

Compound of the present invention have been found in biological tests tobe inhibitors of PI3 kinase. The compounds are selective for class IaPI3 kinases over class Ib and typically exhibit at least a 20-foldselectivity for class Ia over class Ib PI3 kinases. In particular, thecompounds are selective for the p110α isoform.

A compound of the present invention may thus be used as an inhibitor ofPI3 kinase, in particular of a class Ia PI3 kinase. Accordingly, acompound of the present invention can be used to treat a disease ordisorder arising from abnormal cell growth, function or behaviourassociated with PI3 kinase. Examples of such diseases and disorders arediscussed by Drees et al in Expert Opin. Ther. Patents (2004)14(5):703-732. These include cancer, immune disorders, cardiovasculardisease, viral infection, inflammation, metabolism/endocrine disordersand neurological disorders. Examples of metabolism/endocrine disordersinclude diabetes and obesity. Examples of cancers which the presentcompounds can be used to treat include leukaemia, brain tumours, renalcancer, gastric cancer and cancer of the skin, bladder, breast, uterus,lung, colon, prostate, ovary and pancreas. A human or animal patientsuffering from an immune disorder, cancer, cardiovascular disease, viralinfection, inflammation, a metabolism/endocrine disorder or aneurological disorders may thus be treated by a method comprising theadministration thereto of a compound of the present invention as definedabove. The condition of the patient may thereby be improved orameliorated.

In addition to possessing biochemical potency the compounds of theinvention exhibit physicochemical and pharmacokinetic properties whichmake them particularly well adapted for drug use. This is shown forinstance in the results of the biological assays described in Example 3,which follows. In particular the compounds possess high aqueoussolubility at physiological pH; many have a solubility of at least 40 μMand a significant number have a solubility of greater than 100 μM. Highsolubility at physiological pH is desirable since it promotesbioavailability. The compounds also possess high metabolic stability, asshown in particular by the hepatocyte clearance assay described inExample 3 in which most of the tested compounds were shown to have lowhepatocyte clearance. Low hepatocyte clearance correlates with a lowrate of liver metabolism. It can therefore be seen that the compounds ofthe present invention possess improved physicochemical andpharmacokinetic properties whilst retaining biochemical potency asinhibitors of PI3 kinase.

A compound of the present invention can be administered in a variety ofdosage forms, for example orally such as in the form of tablets,capsules, sugar- or film-coated tablets, liquid solutions or suspensionsor parenterally, for example intramuscularly, intravenously orsubcutaneously. The compound may therefore be given by injection orinfusion.

The dosage depends on a variety of factors including the age, weight andcondition of the patient and the route of administration. Daily dosagescan vary within wide limits and will be adjusted to the individualrequirements in each particular case. Typically, however, the dosageadopted for each route of administration when a compound is administeredalone to adult humans is 0.0001 to 50 mg/kg, most commonly in the rangeof 0.001 to 10 mg/kg, body weight, for instance 0.01 to 1 mg/kg. Such adosage may be given, for example, from 1 to 5 times daily. Forintravenous injection a suitable daily dose is from 0.0001 to 1 mg/kgbody weight, preferably from 0.0001 to 0.1 mg/kg body weight. A dailydosage can be administered as a single dosage or according to a divideddose schedule.

A compound is formulated for use as a pharmaceutical or veterinarycomposition also comprising a pharmaceutically or veterinarilyacceptable carrier or diluent. The compositions are typically preparedfollowing conventional methods and are administered in apharmaceutically or veterinarily suitable form. The compound may beadministered in any conventional form, for instance as follows:

A) Orally, for example, as tablets, coated tablets, dragees, troches,lozenges, aqueous or oily suspensions, liquid solutions, dispersiblepowders or granules, emulsions, hard or soft capsules, or syrups orelixirs. Compositions intended for oral use may be prepared according toany method known in the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agentsselected from the group consisting of sweetening agents, flavouringagents, colouring agents and preserving agents in order to providepharmaceutically elegant and palatable preparations.

Tablets contain the active ingredient in admixture with non-toxicpharmaceutically acceptable excipients which are suitable for themanufacture of tablets. These excipients may be for example, inertdiluents, such as calcium carbonate, sodium carbonate, lactose,dextrose, saccharose, cellulose, corn starch, potato starch, calciumphosphate or sodium phosphate; granulating and disintegrating agents,for example, maize starch, alginic acid, alginates or sodium starchglycolate; binding agents, for example starch, gelatin or acacia;lubricating agents, for example silica, magnesium or calcium stearate,stearic acid or talc; effervescing mixtures; dyestuffs, sweeteners,wetting agents such as lecithin, polysorbates or lauryl sulphate. Thetablets may be uncoated or they may be coated by known techniques todelay disintegration and adsorption in the gastrointestinal tract andthereby provide a sustained action over a longer period. For example, atime delay material such as glyceryl monostearate or glyceryl distearatemay be employed. Such preparations may be manufactured in a knownmanner, for example by means of mixing, granulating, tableting, sugarcoating or film coating processes.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is present as such, ormixed with water or an oil medium, for example, peanut oil, liquidparaffin, or olive oil.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example, sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose,sodium alginate, polyvinylpyrrolidone gum tragacanth and gum acacia;dispersing or wetting agents may be naturally-occurring phosphatides,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxyethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides for example polyoxyethylene sorbitan monooleate.

The said aqueous suspensions may also contain one or more preservatives,for example, ethyl or n-propyl p-hydroxybenzoate, one or more colouringagents, such as sucrose or saccharin.

Oily suspension may be formulated by suspending the active ingredient ina vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol.

Sweetening agents, such as those set forth above, and flavouring agentsmay be added to provide a palatable oral preparation. These compositionsmay be preserved by this addition of an antioxidant such as ascorbicacid. Dispersible powders and granules suitable for preparation of anaqueous suspension by the addition of water provide the activeingredient in admixture with a dispersing or wetting agent, a suspendingagent and one or more preservatives. Suitable dispersing or wettingagents and suspending agents are exemplified by those already mentionedabove. Additional excipients, for example sweetening, flavouring andcolouring agents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oils, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally occurring phosphatides, for example soy bean lecithin, andesters or partial esters derived from fatty acids an hexitol anhydrides,for example sorbitan mono-oleate, and condensation products of the saidpartial esters with ethylene oxide, for example polyoxyethylene sorbitanmonooleate. The emulsion may also contain sweetening and flavouringagents. Syrups and elixirs may be formulated with sweetening agents, forexample glycerol, sorbitol or sucrose. In particular a syrup fordiabetic patients can contain as carriers only products, for examplesorbitol, which do not metabolise to glucose or which only metabolise avery small amount to glucose.

Such formulations may also contain a demulcent, a preservative andflavouring and coloring agents;

B) Parenterally, either subcutaneously, or intravenously, orintramuscularly, or intrasternally, or by infusion techniques, in theform of sterile injectable aqueous or oleaginous suspensions. Thissuspension may be formulated according to the known art using thosesuitable dispersing of wetting agents and suspending agents which havebeen mentioned above. The sterile injectable preparation may also be asterile injectable solution or suspension in a non-toxicpaternally-acceptable diluent or solvent, for example as a solution in1,3-butane diol.

Among the acceptable vehicles and solvents that may be employed arewater, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono- or diglycerides. In addition fattyacids such as oleic acid find use in the preparation of injectables;

C) By inhalation, in the form of aerosols or solutions for nebulizers;

D) Rectally, in the form of suppositories prepared by mixing the drugwith a suitable non-irritating excipient which is solid at ordinarytemperature but liquid at the rectal temperature and will therefore meltin the rectum to release the drug. Such materials are cocoa butter andpoly-ethylene glycols;

E) Topically, in the form of creams, ointments, jellies, collyriums,solutions or suspesions.

The invention will be further described in the Examples which follow:

REFERENCE EXAMPLE 1 2,4-Dichloro-thieno[3,2-d]pyrimidine (64)

A mixture of methyl 3-amino-2-thiophenecarboxylate (13.48 g, 85.85 mmol)and urea (29.75 g, 5 eq.) was heated at 190° C. for 2 h. The hotreaction mixture was then poured onto sodium hydroxide solution and anyinsoluble material removed by filtration. The mixture was then acidified(HCl, 2N) to yield 1H-thieno[3,2-d]pyrimidine-2,4-dione (63) as a whiteprecipitate, which was collected by filtration and air dried (9.49 g,66%).

¹H NMR (400 MHz, d₆-DMSO) 6.90 (1H, d, J=5.2 Hz), 8.10 (1H, d, J=5.2Hz), 11.60-11.10 (2H, br s).

A mixture of 1H-thieno[3,2-d]pyrimidine-2,4-dione (9.49 g, 56.49 mmol)and phosphorous oxychloride (150 mL) was heated at reflux for 6 h. Thereaction mixture was then cooled and poured onto ice/water with vigorousstirring yielding a precipitate. The mixture was then filtered to yield2,4-dichloro-thieno[3,2-d]pyrimidine (64) as a white solid (8.68 g, 75%)

¹H NMR (400 MHz, CDCl₃) 7.56 (1H, d, J=5.5 Hz), 8.13 (1H, d, J=5.5 Hz).

REFERENCE EXAMPLE 2 2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(65)

A mixture of 2,4-dichloro-thieno[3,2-d]pyrimidine (64), (8.68 g, 42.34mmol), morpholine (8.11 mL, 2.2 eq.) and MeOH (150 mL) was stirred atroom temperature for 1 h. The reaction mixture was then filtered, washedwith water and MeOH, to yield the title compound as a white solid (11.04g, 100%).

¹H NMR (400 MHz, d₆-DMSO) 3.74 (4H, t, J=4.9 Hz), 3.90 (4H, t, J=4.9Hz), 7.40 (1H, d, J=5.6 Hz), 8.30 (1H, d, J=5.6 Hz).

REFERENCE EXAMPLE 32-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidine-6-carbaldehyde (66)

To a suspension of 2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(65) (1.75 g, 6.85 mmol) in dry THF (40 mL) at −78° C. was added a 2.5Msolution of nBuLi in hexane (3.3 mL, 1.2 eq.). After stirring for 1 h,dry DMF (796 μL, 1.5 eq.) was added. The reaction mixture was stirredfor 1 h at −78° C. and then warmed slowly to room temperature. After afurther 2 h at room temperature the reaction mixture poured ontoice/water yielding a yellow precipitate. This was collected byfiltration and air-dried to yield the title compound (1.50 g, 77%)

¹H NMR (400 MHz, d₆-DMSO) 3.76 (4H, t, J=4.9), 3.95 (4H, t, J=4.9), 8.28(1H, s), 10.20 (1H, s),

REFERENCE EXAMPLE 4 2-Chloro-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine (72)

To a mixture of2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidine-6-carbaldehyde (66)(147 mg, 0.52 mmol), 1-methyl-piperazine (1.5 eq., 87 μL) and aceticacid (1.05 eq., 32 μL) in 1,2-dichloroethane (3 mL) was added sodiumtriacetoxyborohydride (1.1 eq., 121 mg) and then stirred at roomtemperature overnight. The reaction mixture was diluted with DCM, washedwith a saturated solution of sodium hydrogen carbonate, brine, separatedand dried (MgSO₄). The crude product was evaporated in vacuo andpurified by chromatography to give the title compound 72 as an off-whitecrystalline solid (51 mg, 45%).

REFERENCE EXAMPLE 5 Indazole-4-Boronate Ester (70): Route 1

To a solution of 2-methyl-3-nitroaniline (2.27 g, 14.91 mmol) in aceticacid (60 mL) was added a solution of sodium nitrite (1.13 g, 1.1 eq.) inwater (5 mL). After 2 h, the deep red solution was poured onto ice/waterand the resulting precipitate collected by filtration to yield4-nitro-1H-indazole (67) (1.98 g, 81%).

A mixture of 4-nitro-1H-indazole (760 mg, 4.68 mmol), palladium oncharcoal (10%, cat.) and ethanol (30 mL) was stirred under a balloon ofhydrogen for 4 h. The reaction mixture was then filtered through celite,and the solvent removed in vacuo to yield 1H-indazol-4-ylamine (68) (631mg, 100%).

An aqueous solution of sodium nitrite (337 mg, 4.89 mmol) in water (2mL) was added dropwise to a suspension of 1H-indazol-4-ylamine (631 mg,4.74 mmol) in 6M hydrochloric acid (7.2 mL) at below 0° C. Afterstirring for 30 minutes, sodium tetrafluorobrate (724 mg) was added tothe reaction mixture. A viscous solution resulted, which was filteredand washed briefly with water to yield 1H-indazole-4-diazoniumtetrafluoroborate salt (69) (218 mg, 20%) as a deep red solid.

Dry MeOH (4 mL) was purged with argon for 5 minutes. To this was added1H-indazole-4-diazonium tetrafluoroborate salt (218 mg, 0.94 mmol),bis-pinacolato diboron (239 mg, 1.0 eq.) and[1,1′-bis(diphenylphosphino)ferrocene]palladium (II) chloride (20 mg).The reaction mixture was stirred for 5 h and then filtered throughcelite. The residue was purified using flash chromatography to yield thedesired title compound (70), (117 mg).

REFERENCE EXAMPLE 6 Indazole-4-Boronate Ester (70): Route 2

To a solution of 3-bromo-2-methyl aniline (5.0 g, 26.9 mmol) inchloroform (50 mL) was added potassium acetate (1.05 eq., 28.2 mmol,2.77 g). Acetic anhydride (2.0 eq., 53.7 mmol, 5.07 mL) was added withconcurrent cooling in ice-water. The mixture was then stirred at roomtemperature for 10 minutes after which time a white gelatinous solidformed. 18-Crown-6 (0.2 eq., 5.37 mmol, 1.42 g) was then added followedby iso-amyl nitrite (2.2 eq., 59.1 mmol, 7.94 mL) and the mixture washeated under reflux for 18 h. The reaction mixture was allowed to cool,was partitioned between chloroform (3×100 mL) and saturated aqueoussodium hydrogen carbonate (100 mL). The combined organic extracts werewashed with brine (100 mL), separated and dried (MgSO₄).

The crude product was evaporated onto silica and purified bychromatography eluting with 20%→40% EtOAc-petrol to give the N-acetylindazole (73) (3.14 g, 49%) as an orange solid followed by the indazole(74) (2.13 g, 40%) as a pale orange solid.

73: ¹H NMR (400 MHz, CDCl₃) 2.80 (3H, s), 7.41 (1H, t, J=7.8 Hz), 7.50(1H, d, J=7.8 Hz), 8.15 (1H, s), 8.40 (1H, d, J=7.8 Hz).

74: ¹H NMR (400 MHz, CDCl₃) 7.25 (1H, t, J=7.3 Hz), 7.33 (1H, d, J=7.3Hz), 7.46 (1H, d, J=7.3 Hz), 8.11 (1H, s), 10.20 (1H, br s),

To a solution of the N-acetyl indazole (3.09 g, 12.9 mmol) in MeOH (50mL) was added 6N aqueous HCl (30 mL) and the mixture was stirred at roomtemperature for 7 h. The MeOH was evaporated and the mixture partitionedbetween EtOAc (2×50 mL) and water (50 mL). The combined organic layerswere washed with brine (50 mL), separated and dried (MgSO₄). The solventwas removed by evaporation under reduced pressure to give an orangesolid (2.36 g, 93%).

To a solution of the 4-bromoindazole (500 mg, 2.54 mmol) andbis(pinacolato)diboron (1.5 eq., 3.81 mmol) in DMSO (20 mL) was addedpotassium acetate (3.0 eq., 7.61 mmol, 747 mg; dried in drying pistol)and PdCl₂(dppf)₂ (3 mol %, 0.076 mmol, 62 mg). The mixture was degassedwith argon and heated at 80° C. for 40 h. The reaction mixture wasallowed to cool and partitioned between water (50 mL) and ether (3×50mL). The combined organic layers were washed with brine (50 mL),separated and dried (MgSO₄). The crude material was purified bychromatography eluting with 30%→40% EtOAc-petrol to give an inseparable3:1 mixture of the boronate ester (369 mg, 60%) and indazole (60 mg,20%); this was isolated as a yellow gum which solidified upon standingto furnish (70) as an off-white solid.

¹H NMR (400 MHz, d₆-DMSO) (70) 1.41 (12H, s), 7.40 (1H, dd, J=8.4 Hz,6.9 Hz), 7.59 (1H, d, J=8.4 Hz), 7.67 (1H, d, J=6.9 Hz), 10.00 (1H, brs), 8.45 (1H, s), and indazole: 7.40 (1H, t), 7.18 (1H, t, J=7.9 Hz),7.50 (1H, d, J=9.1 Hz), 7.77 (1H, d, J=7.9 Hz), 8.09 (1H, s). Impurityat 1.25.

REFERENCE EXAMPLE 7 6-Fluoroindazole-4-Boronate Ester (75)

To a solution of 4-fluoro-2-nitrotoluene (3.44 g) in trifluoroaceticacid (13 mL) was added concentrated sulfuric acid (4 mL) followed byN-bromosuccinimide (5.92 g). The reaction mixture was stirred for 16 h.and was then quenched with brine, extracted into ethyl acetate, anddried (MgSO₄). The solvent was removed in vacuo to furnish crude1-bromo-5-fluoro-2-methyl-3-nitro-benzene (5.96 g).

To a solution of crude 1-bromo-5-fluoro-2-methyl-3-nitro-benzene (5.96g) in MeOH (90 mL) was added concentrated hydrochloric acid (11.7 mL)and iron (6.1 g) and the reaction mixture was heated to reflux. After 16h, the mixture was cooled, diluted with DCM, washed with sodiumcarbonate solution, dried (MgSO₄) and the solvent removed in vacuo. Theresidue was purified using flash chromatography to yield3-bromo-5-fluoro-2-methyl-phenylamine (1.46 g).

To a solution of 3-bromo-5-fluoro-2-methyl-phenylamine (470 mg) indioxane (6 mL) was added triethylamine (1.28 mL), palladium acetate (25mg), 2-dicyclohexylphosphino biphenyl (161 mg) and pinacol borane (1.001ml) and the mixture was heated to 80° C. for 4 h. The reaction mixturewas cooled, diluted with chloroform, washed with brine, dried (MgSO₄)and the solvent removed in vacuo. The residue was purified using flashchromatography to yield the desired title compound (466 mg).

REFERENCE EXAMPLE 8 Preparation of2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine-6-carbaldehyde(71)

A mixture of2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidine-6-carbaldehyde (66)(100 mg, 0.35 mmol),4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indazole (70) (95mg, 0.39 mmol) and sodium carbonate (112 mg) were suspended in toluene(2.5 mL), ethanol (1.5 mL) and water (0.7 mL). To this was addedbis(triphenylphosphine)palladium(II) chloride (13.5 mg) and the reactionvessel was flushed with argon. The reaction mixture was microwaved at120° C. for 1 h and then partitioned between DCM and water, the organiclayer was washed with brine, dried over magnesium sulfate, filtered andevaporated in vacuo. The resulting residue was purified using flashchromatography to yield the title compound 71 (97 mg).

EXAMPLE 12-(1H-Indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(59) by Route 1

To a mixture of2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine-6-carbaldehyde(91 mg, 0.26 mmol), 1-methylpiperazine (34 mg, 0.36 mmol) and aceticacid (15 uL) in 1,2-dichloroethane (2 mL) was added sodiumtriacetoxyborohydride (60 mg, 0.28 mmol). The reaction mixture wasstirred at room temperature overnight and then basified (NaHCO₃,saturated), diluted with DCM, and washed with brine. The organic layerwas separated, dried (MgSO₄), filtered and evaporated in vacuo. Theresidue was purified using flash chromatography to give the titlecompound (33 mg).

¹H NMR 400 MHz, d₆-DMSO) 2.18 (s, 3H), 2.30-2.45 (br m, 4H), 2.48-2.55(br m, 4H), 3.82-3.84 (m, 4H), 3.86 (s, 2H), 3.98-4.00 (m, 4H),7.44-7.47 (m, 2H), 7.65 (d, 1H, J=8.2 Hz), 8.21 (d, 1H, J=7.2 Hz), 8.87(s, 1H), 13.16 (br s, 1H); MS (ESI⁺) 450.1 (MH⁺).

EXAMPLE 22-(1H-Indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(59) by Route 2

Indazole-4-boronate ester (2.0 eq., 0.82 mmol),2-Chloro-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(compound 72 prepared in Reference Example 4: 150 mg, 0.41 mmol) andsodium carbonate (3.0 eq., 130 mg) were combined in a mixture of toluene(2 mL), ethanol (1 mL) and water (0.5 mL). PdCl₂(PPh₃)₂ (0.1 eq.) wasadded and the reaction mixture was flushed with Argon, and then heatedin a microwave reactor at 130° C. for 2 h. Following extraction(DCM/brine) and flash column chromatography, the product was isolated asa white solid (149 mg, 81%)

EXAMPLE 2A Further Compounds of the Invention

The following compounds of the invention were prepared by analogy withthe process of Example 2. Compound 72 was replaced in each case by theappropriate precursor chloro compound, prepared by the method ofReference Example 4 using the relevant amine in place of 1-methylpiperazine. The preparation of the amine is described below wherenecessary. NMR data are given for each of the title compounds of theinvention.

2-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-ethanol(60)

Prepared via2-[4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazin-1-yl]-ethanol.

¹H NMR 400 MHz, d₆-DMSO) 2.40 (br m, 2H), 2.42-2.52 (b, 8H, under DMSOpeak), 3.48 (q, 2H, J=6.0 Hz), 3.82-3.86 (m, 6H), 3.98-4.01 (m, 4H),4.34 (br s, 1H,), 7.44-7.48 (m, 2H), 7.65 (d, 1H, J=8.3 Hz), 8.21 (d,1H, J=6.8 Hz), 8.87 (s, 1H), 13.15 (br s, 1H); MS (ESI⁺) 480.1 (MH⁺).

4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-sulfonicacid dimethylamide (62)

Prepared Via4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazine-1-sulfonicacid dimethylamide.

¹H NMR 400 MHz, CDCl₃) 2.63-2.66 (m, 4H), 2.84 (s, 6H), 3.31-3.34 (m,4H), 3.89 (s 2H), 3.92-3.94 (m, 4H), 4.08-4.11 (m, 4H), 7.39 (s, 1H),7.51 (t, 1H, J=8.1 Hz), 7.60 (d, 1H, J=8.1 Hz), 8.28 (d, 1H, J=6.7 Hz),9.02 (s, 1H), 10.12 (br s, 1H); MS (ESI⁺) 543.1 (MH⁺)

{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-morpholin-4-yl-methanone(76)

Prepared via[4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazin-1-yl]-morpholin-4-yl-methanone,prepared from morpholin-4-yl-piperazin-1-yl-methanone.

Amine preparation: a mixture of 4-morpholinocarbonyl chloride (0.38 ml),1-BOC-piperazine (552 mg) and potassium carbonate (439 mg) in MeCN (7mL) was stirred at room temperature for 3 h. The reaction mixture wasthen diluted with DCM, washed with brine, dried (MgSO₄) and the solventremoved in vacuo to yield4-(morpholine-4-carbonyl)-piperazine-1-carboxylic acid tert-butyl ester(865 mg). Treatment of this compound with HCl in. DCM/MeOH yielded thedesired compound, which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 2.55-2.58 (4H, m), 3.28-3.32 (4H, m), 3.35-3.39(4H, m), 3.67-3.71 (4H, m), 3.88 (2H, s), 3.92-3.96 (4H, m), 4.08-4.12(4H, m), 7.39 (1H, s), 7.52 (1H, t, J=8.0 Hz), 7.60 (1H, d, J=6.3 Hz),8.30 (1H, d, J=7.0 Hz), 9.02 (1H, s), 10.10 (1H, br); MS (ESI⁺) 549(MH⁺).

4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-carboxylicacid (2-methoxy-ethyl)-methyl-amide (77)

Prepared via4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazine-1-carboxylicacid (2-methoxy-ethyl)-methyl-amide, prepared frompiperazine-1-carboxylic acid (2-methoxy-ethyl)-methyl-amide.

Amine preparation: to N—BOC-piperazine (500 mg) in DCM (5 mL) andtriethylamine (0.41 ml) was added 4-nitrophenyl chloroformate (541 mg).After 1 h the reaction mixture was diluted with DCM, washed with brine,dried (MgSO₄) and the solvent removed in vacuo to yieldpiperazine-1,4-dicarboxylic acid tert-butyl ester 4-nitro-phenyl ester(940 mg).

To piperazine-1,4-dicarboxylic acid tert-butyl ester 4-nitro-phenylester (500 mg) in THF (5 mL) was added N-(2-methoxyethyl)methylamine(254 mg) and the reaction mixture was heated to reflux for 24 h. Thereaction mixture was concentrated in vacuo and purified using flashcolumn chromatography to yield4-[(2-methoxy-ethyl)-methyl-carbamoyl]-piperazine-1-carboxylic acidtert-butyl ester (304 mg). Treatment of this compound with HCl inDCM/MeOH yielded the desired compound, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 2.59-2.63 (4H, m), 2.90 (3H, s), 3.27-3.30 (4H,m), 3.31 (3H, s), 3.48 (2H, t), 3.57 (2H, t), 3.90 (2H, s), 3.92-3.96(4H, m), 4.08-4.12 (4H, m), 7.39 (1H, s), 7.52 (1H, t), 7.60 (1H, d,J=6.3 Hz), 8.30 (1H, d, J=7.0 Hz), 9.02 (1H, s), 10.10 (1H, br); MS(ESI⁺) 551 (MH⁺).

2-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-N,N-dimethyl-acetamide(78)

Prepared via2-[4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazin-1-yl]-N,N-dimethylacetamide,prepared from N,N-dimethyl-2-piperazin-1-yl-acetamide.

Amine preparation: a mixture of 1-BOC-piperazine (387 mg),2-chloro-N,N-dimethylacetamide (0.43 mL) and triethylamine (0.58 mL) inchloroform was stirred at room temperature. After stirring overnight thereaction mixture was diluted with DCM, washed with brine, dried (MgSO₄)and the solvent removed in vacuo to yield4-dimethylcarbamoylmethyl-piperazine-1-carboxylic acid tert-butyl ester(558 mg). Treatment of this compound with HCl in DCM/MeOH yielded thedesired compound, which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 2.63 (sbr, 8H, 4×CH₂), 2.95 (s, 3H, CH₃), 3.07(s, 3H, CH₃), 3.20 (s, 2H, CH₂), 3.85 (s, 2H, CH₂), 3.90-3.93 (m, 4H,2×CH₂), 4.07-4.10 (m, 4H, 2×CH₂), 4.36 (s, H, ArH), 7.49 (t, H, ArH,J=7.74 Hz), 7.57 (d, H, ArH, J=8.26 Hz), 8.26 (d, H, ArH, J=7.23 Hz),9.00 (s, H, ArH), 10.25 (br s, H, NH); MS (ESI⁺) 521.29 (MH⁺).

4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-carboxylicacid dimethylamide (79)

Prepared via4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazine-1-carboxylicacid dimethylamide, prepared from piperazine-1-carboxylic aciddimethylamide.

Amine preparation: To a solution of 1-BOC-piperazine (867 mg) in dry THF(8 mL) was added triethylamine (0.97 mL) followed by dimethylcarbamoylchloride (0.51 mL). After stirring for 24 h the reaction mixture wasthen diluted with DCM, washed with brine, dried (MgSO₄) and the solventwas removed in vacuo to yield4-dimethylcarbamoyl-piperazine-1-carboxylic acid tert-butyl ester (940mg). Treatment of this compound with HCl in DCM/MeOH yielded the desiredcompound, which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 2.57-2.61 (4H, m), 2.87 (6H, s), 3.30-3.35 (4H,m), 3.89 (2H, s), 3.92-3.96 (4H, m), 4.08-4.12 (4H, m), 7.39 (1H, s),7.52 (1H, t, J=8.0 Hz), 7.60 (1H, d, J=6.3), 8.30 (1H, d, J=7.0 Hz),9.02 (1H, s), 10.10 (1H, br); (ESI+): MS (ESI⁺) 507 (MH⁺).

2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(3-morpholin-4-yl-propane-1-sulfonyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine(80)

Prepared via2-Chloro-4-morpholin-4-yl-6-[4-(3-morpholin-4-yl-propane-1-sulfonyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine,prepared from 4-[3-(piperazine-1-sulfonyl)-propyl]-morpholine.

Amine preparation: A mixture of 1-BOC-piperazine (3.26 g),3-chloropropanesulfonyl chloride (2.63 g) and triethylamine (2.68 mL)was stirred at room temperature in DCM (25 mL). After 2 h the reactionmixture was diluted with DCM, washed with brine, dried (MgSO₄) and thesolvent removed in vacuo to yield4-(3-chloro-propane-1-sulfonyl)-piperazine-1-carboxylic acid tert-butylester (4.65 g). A mixture of4-(3-chloro-propane-1-sulfonyl)-piperazine-1-carboxylic acid tert-butylester (4.65 g), potassium iodide (1.1 g), potassium carbonate andmorpholine (1.6 mL) was heated to reflux in MeCN (100 ml). After 16 h,the reaction mixture was cooled, diluted with DCM, washed with brine,dried (MgSO₄) and the solvent removed in vacuo to yield4-(3-morpholin-4-yl-propane-1-sulfonyl)-piperazine-1-carboxylic acidtert-butyl ester (4.8 g). Treatment of this compound with HCl inDCM/MeOH yielded the desired compound, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 1.98-2.02 (m, 2H), 2.44-2.47 (m, 6H), 2.67-2.69(m, 4H), 2.99-3.03 (m, 2H), 3.36-3.38 (m, 4H), 3.69-3.71 (m, 4H), 3.90(s, 2H), 3.91-3.93 (m, 4H), 4.08-4.10 (m, 4H), 7.39 (s, H, ArH), 7.50(t, H, ArH, J=7.7 Hz), 7.58 (d, H, ArH, J=8.32 Hz), 8.27 (d, H, ArH,J=7.44 Hz), 9.00 (s, H, ArH), 10.10 (br s, H, NH); MS (ESI⁺) 627.29(MH⁺).

{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-methyl-amine(81)

Prepared via[1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-(2-methoxy-ethyl)-methyl-amine,prepared from (2-methoxy-ethyl)-methyl-piperidin-4-yl-amine.

Amine preparation: a mixture of N-BOC-4-piperidine (500 mg),N-(2-methoxyethyl)methylamine (335 mg), acetic acid (0.15 mL) and sodiumtriacetoxyborohydride (797 mg) was stirred at room temperature in1,2-dichloroethane (5 mL). After stirring overnight, the reactionmixture was diluted with chloroform, washed with sodium bicarbonatesolution, dried (MgSO₄) and the solvent removed in vacuo. The residuewas purified using flash column chromatography to yield4-[(2-methoxy-ethyl)-methyl-amino]-piperidine-1-carboxylic acidtert-butyl ester. Treatment of this compound with HCl in DCM/MeOHyielded the desired compound, which was isolated as the hydrochloridesalt.

¹H NMR (400 MHz, CDCl₃): 1.62-1.72 (2H, m), 1.76-1.84 (2H, m), 2.10-2.18(2H, m), 2.36 (3H, s), 2.40-2.48 (1H, m), 2.68 (2H, t, J=6.0 Hz),3.04-3.11 (2H, m), 3.38 (3H, s), 3.50 (2H, t, J=6.3 Hz), 3.85 (2H, s),3.92-3.97 (4H, m), 4.08-4.12 (4H, m), 7.39 (1H, s), 7.52 (1H, t, J=8.0Hz), 7.60 (1H, d, J=6.3 Hz), 8.30 (1H, d, J=7.0 Hz), 9.02 (1H, s), 10.10(1H, br); MS (ESI⁺) 522 (MH⁺).

3-4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-sulfonyl}-propyl)-dimethyl-amine(82)

Prepared via{3-[4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazine-1-sulfonyl]-propyl}-dimethyl-amine,prepared from dimethyl-[3-(piperazine-1-sulfonyl)-propyl]-amine. Theamine preparation was as for compound 80 above.

¹H NMR (400 MHz, CDCl₃): 2.00-2.08 (2H, m), 2.26 (6H, s), 2.42 (2H, t,J=6.7), 2.68-2.72 (4H, m), 3.00-3.05 (2H, m), 3.37-3.41 (4H, m), 3.90(2H, s), 3.92-3.96 (4H, m), 4.08-4.12 (4H, m), 7.39 (1H, s), 7.52 (1H,t, J=8.0), 7.60 (1H, d, J=6.3), 8.30 (1H, d, J=7.0), 9.02 (1H, s), 10.10(1H, br); MS (ESI⁺) 585 (MH⁺).

2-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-2-methyl-propan-1-ol(83)

Prepared via2-[4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazin-1-yl]-2-methyl-propan-1-ol,prepared from 2-methyl-2-piperazin-1-yl-propan-1-ol.

Amine preparation: a mixture of BOC-piperazine (1.87 g), ethyl2-bromoisobutyrate (5.90 g) and potassium carbonate (1.53 g) in MeCN (20mL) was heated in a sealed tube at 80° C. for 3 days. The reactionmixture was cooled, diluted with chloroform, washed with brine, dried(MgSO₄) and the solvent removed in vacuo. The residue was purified byflash column chromatography to yield4-(1-ethoxycarbonyl-1-methyl-ethyl)-piperazine-1-carboxylic acidtert-butyl ester (2.97 g).

Treatment of 4-(1-ethoxycarbonyl-1-methyl-ethyl)-piperazine-1-carboxylicacid tert-butyl ester with lithium aluminium hydride in ether yieldedthe corresponding alcohol,4-(2-hydroxy-1,1-dimethyl-ethyl)-piperazine-1-carboxylic acid tert-butylester. Treatment of this compound with HCl in DCM/MeOH yielded thedesired compound, which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 9.02 (1H, s), 8.25 (1H, d), 7.60 (1H, d),7.51-7.49 (1H, m), 7.39 (1H, s), 4.08-4.06 (4H, m), 3.90-3.88 (4H, m),3.85 (2H, s), 3.46 (2H, s), 2.70-2.50 (8H, m), 1.05 (6H, s); MS (ESI⁺)508 (MH⁺).

1′-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-[1,4′]bipiperidinyl(84)

Prepared via1′-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-[1,4′]bipiperidinyl,prepared from 4-piperidinopiperidine.

¹H NMR (400 MHz, CDCl₃): 1.40-1.50 (2H, br), 1.50-1.75 (6H, br),1.80-1.90 (2H, br), 2.21 (2H, t, J=10.7), 2.35-2.43 (1H, br), 2.50-2.60(4H, br), 3.04-3.10 (2H, br. d, J=11.4), 3.84 (2H, s), 3.92-3.96 (4H,m), 4.08-4.12 (4H, m), 7.39 (1H, s), 7.52 (1H, t, J=8.0), 7.60 (1H, d,J=6.3), 8.30 (1H, d, J=7.0), 9.02 (1H, s), 10.10 (1H, br); MS (ESI⁺) 518(MH⁺).

2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-morpholin-4-yl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine(85)

Prepared via2-Chloro-4-morpholin-4-yl-6-(4-morpholin-4-yl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared from 4-morpholinopiperidine.

¹H NMR (400 MHz, CDCl₃): 1.55-1.68 (2H, m), 1.83-1.90 (2H, m), 2.11-2.18(2H, m), 2.18-2.25 (1H, m), 2.54-2.60 (4H, m), 3.05-3.11 (2H, m),3.70-3.76 (4H, m), 3.84 (2H, s), 3.92-3.96 (4H, m), 4.08-4.12 (4H, m),7.39 (1H, s), 7.52 (1H, t, J=8.0), 7.60 (1H, d, J=6.3), 8.30 (1H, d,J=7.0), 9.02 (1H, s), 10.10 (1H, br); MS (ESI⁺) 520 (MH⁺).

2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyrimidin-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine(86)

Prepared via2-Chloro-4-morpholin-4-yl-6-(4-pyrimidin-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared from 1-(2-pyrimidyl)piperazine.

¹H NMR (400 MHz, CDCl₃): 2.64-2.70 (4H, m), 3.87-3.96 (10H, m),4.10-4.14 (4H, m), 6.50 (1H, t, J=4.8), 7.40 (1H, s), 7.52 (1H, t,J=7.8), 7.60 (1H, d, J=8.3), 8.29-8.33 (3H, m), 9.02 (1H, s), 10.10 (1H,br); MS (ESI⁺) 514 (MH⁺).

1-(2-Hydroxy-ethyl)-4-[2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-2-one(87)

Prepared via4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-1-(2-hydroxy-ethyl)-piperazin-2-one,prepared from 1-(2-hydroxy-ethyl)-piperazin-2-one.

Amine preparation: to 4-CBZ-piperazin-2-one (1.95 g) in DMF (5 mL) at 0°C. was added sodium hydride (60% dispersion in mineral oil, 660 mg) inseveral aliquots. After stirring for 1 h, 2-bromoethylacetate (1.38 ml)was added. The reaction mixture was stirred at room temperatureovernight; it was then diluted with ethyl acetate, washed with brine,dried (MgSO₄) and the solvent was removed in vacuo. The resultingresidue was purified using flash chromatography to yield4-(2-acetoxy-ethyl)-3-oxo-piperazine-1-carboxylic acid benzyl ester (925mg).

¹H NMR (400 MHz, d₆-DMSO): 2.77 (2H, d, J=5.5 Hz), 3.16 (2H, s),3.32-3.36 (2H, m), 3.38-3.42 (2H, m), 3.51-3.55 (2H, m), 3.80-3.85 (4H,m), 3.97 (2H, s), 4.00-4.04 (4H, m), 4.70 (1H, t, J=5.4 Hz, OH), 7.45(1H, t, J=7.7 Hz), 7.50 (1H, s), 7.66 (1H, d, J=8.2 Hz), 8.22 (1H, d,J=7.3 Hz), 8.89 (1H, s), 13.15 (1H, br, NH); MS (ESI⁺) 494 (MH⁺).

6-(4-Cyclopropylmethyl-piperazin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(88)

Prepared via2-Chloro-6-(4-cyclopropylmethyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-cyclopropylmethyl-piperazine.

Amine preparation: A mixture of BOC-piperazine (887 mgg),(bromomethyl)cyclopropane (0.5 mL) and potassium carbonate (779 mg) inMeCN (10 mL) was heated to reflux for 16 h. The reaction mixture wascooled, diluted with chloroform, washed with brine, dried (MgSO₄) andthe solvent removed in vacuo. The residue was purified using flashchromatography to yield 4-yclopropylmethyl-piperazine-1-carboxylic acidtert-butyl ester (1.05 g). Treatment of this compound with HCl inDCM/MeOH yielded the desired compound, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 0.07-0.14 (m, 2H, 2×CH₂), 0.48-0.51 (m, 2H,2×CH), 0.8-0.95 (m, H, CH), 2.28-2.32 (m, 2H, CH₂), 2.5-2.7 (m, 8H,4×CH₂), 3.86 (s, 2H, CH₂), 3.90-3.93 (m, 4H, 2×CH₂), 4.07-4.11 (m, 4H,2×CH₂), 7.38 (s, H, ArH), 7.50 (t, H, ArH, J=7.79 Hz), 7.58 (d, H, ArH,J=8.28 Hz), 8.28 (d, H, ArH, J=7.57 Hz), 9.02 (s, H, ArH), 10.15 (sbr,H, NH); MS (ESI⁺) 490.19 (MH⁺).

2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine(89)

Prepared via2-Chloro-4-morpholin-4-yl-6-(4-pyridin-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared from 1-pyridin-2-yl-piperazine (commercially available).

¹H NMR (400 MHz, CDCl₃): 10.1 (1H, br s), 9.02 (1H, s), 8.25 (1H, d),8.22-8.20 (1H, m), 7.60 (1H, d), 7.51-7.43 (2H, m), 7.39 (1H, s),6.61-6.60 (1H, m), 4.08-4.06 (4H, m), 3.90-3.88 (6H, m), 3.60-3.58 (4H,m), 2.72-2.70 (4H, m); MS (ESI⁺) 513 (MH⁺).

2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine(90)

Prepared via2-Chloro-4-morpholin-4-yl-6-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine,prepared from 1-(2,2,2-trifluoro-ethyl)-piperazine.

Amine preparation: to BOC-piperazine (4 g) in DCM (40 mL) was addedtrifluoroacetic anhydride (6.06 mL) and triethylamine (3.29 mL). Afterstirring overnight the reaction mixture was diluted with diluted withDCM, washed with sodium bicarbonate solution, dried (MgSO₄) and thesolvent removed in vacuo to yield4-(2,2,2-trifluoro-acetyl)-piperazine-1-carboxylic acid tert-butyl ester(6.06 g).

To 4-(2,2,2-trifluoro-acetyl)-piperazine-1-carboxylic acid tert-butylester (6.06 g) in dry THF (60 mL) was added borane dimethyl sulfidecomplex (4.5 ml) and the reaction mixture was heated to reflux. After 2h the reaction mixture was cooled to 0° C. and MeOH was carefully added,followed by water. The organics were extracted into ethyl acetate, dried(MgSO₄) and the solvent removed in vacuo to yield4-(2,2,2-trifluoro-ethyl)-piperazine-1-carboxylic acid tert-butyl ester(4.46 g). Treatment with HCl in DCM/MeOH yielded the desired compound,which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 2.56 (4H, m), 2.69 (4H, m), 2.93 (2H, q), 3.79(2H, s), 3.85 (4H, m), 4.02 (4H, m), 7.23 (1H, s), 7.44 (1H, d), 7.52(1H, d), 8.21 (1H, d), 8.94 (1H, s).

2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-thiazol-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine(91)

Prepared via2-Chloro-4-morpholin-4-yl-6-(4-thiazol-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared from 1-thiazol-2-yl-piperazine (commercially available).

¹H NMR (400 MHz, CDCl₃): 10.1 (1H, br s), 9.02 (1H, s), 8.25 (1H, d),7.60 (1H, d), 7.51-7.49 (2H, m), 7.39 (1H, s), 7.20 (1H, d), 6.60 (1H,d), 4.08-4.06 (4H, m), 3.90-3.88 (6H, m), 3.55-3.50 (4H, m), 2.72-2.70(4H, m); MS (ESI⁺) 519 (MH⁺).

2-(6-Fluoro-1H-indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(92)

Prepared by treatment of5-fluoro-2-methyl-3-[6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-2-yl]-phenylaminewith isoamyl nitrite in chloroform and acetic acid.

¹H NMR (400 MHz, CDCl₃): 10.1 (1H, br s), 9.02 (1H, s), 8.10 (1H, dd),7.39 (1H, s), 7.22 (1H, dd), 4.08-4.06 (4H, m), 3.90-3.88 (4H, m), 3.85(2H, s), 2.70-2.50 (8H, m), 2.30 (3H, s); MS (ESI⁺) 468 (MH⁺).

2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-2-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine(93)

Prepared via2-Chloro-4-morpholin-4-yl-6-(4-pyridin-2-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine.The amine was prepared as described for compound 95.

¹H NMR (400 MHz, CDCl₃): 2.61 (m, 8H, 4×CH₂), 3.70 (s, 2H, CH₂), 3.86(s, 2H, CH₂), 3.90-3.93 (m, 4H, 2×CH₂), 4.07-4.10 (m, 4H, 2×CH₂),7.14-7.17 (m, H, ArH), 7.36 (s, H, ArH), 7.40 (d, H, ArH, J=7.78 Hz),7.49 (t, H, ArH, J=7.77 Hz), 7.57 (d, H, ArH, J=8 Hz), 7.64 (t, H, ArH,J=7.64 Hz), 8.27 (d, H, ArH, J=6.64 Hz), 8.56 (d, H, ArH, J=4.83 Hz),9.0 (s, H, ArH), 10.12 (sbr, H, NH); MS (ESI⁺) 527.28 (MH⁺).

2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-thiazol-2-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine(94)

Prepared via2-Chloro-4-morpholin-4-yl-6-(4-thiazol-2-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared from 1-thiazol-2-ylmethyl-piperazine. The amine was prepared asdescribed for compound 95 below.

¹H NMR (400 MHz, CDCl₃): 2.67 (m, 8H, 4×CH₂), 3.87 (s, 2H, CH₂),3.91-3.93 (m, 6H, 3×CH₂), 4.07-4.10 (m, 4H, 2×CH₂), 7.28 (d, H, ArH,J=3.23 Hz), 7.37 (s, H, ArH), 7.49 (t, H, ArH, J=7.73 Hz), 7.58 (d, H,ArH, J=8.31 Hz), 7.70 (d, H, ArH, J=3.32 Hz), 8.27 (d, H, ArH, J=6.79Hz), 9.0 (s, H, ArH), 10.1 (sbr, H, NH).

2-(1H-Indazol-4-yl)-6-[4-(5-methyl-furan-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(95)

Prepared via2-Chloro-6-[4-(5-methyl-furan-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-(5-methyl-furan-2-ylmethyl)-piperazine.

Amine preparation: A mixture of 1-BOC-piperazine (1.63 g), 5-methylfurfural (964 mg) and acetic acid (0.50 mL) was stirred in1,2-dichloroethane (10 mL) at room temperature. To this was added sodiumtriacetoxyborohydride (2.04 g) and the reaction mixture was stirredovernight. The reaction mixture was diluted with chloroform, washed withbrine, dried (MgSO₄) and the solvent was removed in vacuo to liberate4-(5-methyl-furan-2-ylmethyl)-piperazine-1-carboxylic acid tert-butylester as an orange oil. Treatment of this compound with HCl in DCM/MeOHyielded the desired compound, which was isolated as the hydrochloridesalt.

¹H NMR (400 MHz, CDCl₃): 10.1 (1H, br s), 9.02 (1H, s), 8.28 (1H, d),7.60 (1H, d), 7.51-7.48 (1H, m), 6.10 (1H, d), 5.88 (1H, d), 4.08-4.06(4H, m), 3.90-3.88 (4H, m), 3.83 (2H, s), 3.51 (2H, s), 2.70-2.50 (8H,m), 2.26 (3H, s); MS (ESI⁺) 530 (MH⁺).

1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylicacid amide (96)

Prepared via1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidine-4-carboxylicacid amide, prepared using isonipecotamide (commercially available).

¹H NMR (400 MHz, d₆-DMSO): 1.52-1.74 (4H, m); 2.00-2.16 (3H, m);2.90-2.98 (2H, m); 3.80-3.90 (6H, m); (4H, t, J=4.7 Hz); 6.70 (1H, s);7.20 (1H, s); 7.48 (2H, t, J=7.7 Hz); 7.65 (1H, d, J=8.2 Hz); 8.22 (1H,d, J=7.3 Hz), 8.88 (1H, s), 13.15 (1H, s); MS (ESI⁺) 478 (MH⁺).

2-(1H-Indazol-4-yl)-6-[4-(2-methoxy-1,1-dimethyl-ethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(97)

Prepared via2-Chloro-6-[4-(2-methoxy-1,1-dimethyl-ethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-(2-methoxy-1,1-dimethyl-ethyl)-piperazine.

Amine preparation: a mixture of benzylpiperazine, methoxyacetyl chlorideand triethylamine was stirred in DCM for 2 h to yield1-(4-benzyl-piperazin-1-yl)-2-methoxy-ethanone following standard workup.

To a solution of 1-(4-benzyl-piperazin-1-yl)-2-methoxy-ethanone (6.14 g)in dry THF (80 mL) at −10° C. was added zirconium chloride (5.76 g).After 30 minutes methyl magnesium bromide (3.0M solution in ether, 49.6mL) was added in a dropwise manner. The reaction mixture was allowed towarm to room temperature. After stirring for 1 day, the reaction mixturewas cooled, diluted with chloroform, washed with brine, dried (MgSO₄)and the was solvent removed in vacuo. The resulting residue was purifiedusing flash chromatography to yield1-benzyl-4-(2-methoxy-1,1-dimethyl-ethyl)-piperazine. Subsequenttransfer hydrogenation, using ammonium formate and 10% palladium oncarbon in MeOH, furnished the desired compound.

¹H NMR (400 MHz, CDCl₃): 1.07 (s, 6H, 2×CH₃), 2.61 (m, 4H, 2×CH₂), 2.69(m, 4H, 2×CH₂), 3.26 (s, 2H, CH₂), 3.33 (s, 3H, CH₃), 3.83 (s, 2H, CH₂),3.90-3.93 (m, 4H, 2×CH₂), 4.07-4.10 (m, 4H, 2×CH₂), 7.36 (s, H, ArH),7.49 (t, H, ArH, J=7.72 Hz), 7.57 (d, H, ArH, J=8.25 Hz), 8.26 (d, H,ArH, J=7.13 Hz), 9.0 (s, H, ArH), 10.1 (sbr, H, NH); MS (ESI⁺) 522(MH⁺).

2-(1H-Indazol-4-yl)-6-[(3R,5S)-4-(2-methoxy-ethyl)-3,5-dimethyl-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(98)

Prepared via2-Chloro-6-[(3R,5S)-4-(2-methoxy-ethyl)-3,5-dimethyl-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from (2R,6S)-1-(2-methoxy-ethyl)-2,6-dimethyl-piperazine.

Amine preparation: to a solution of 2,6-dimethylpiperazine(predominantly cis) (250 mg), tert-butanol (2.5 mL), sodium hydroxide(88 mg) and water (0.5 mL) was added a solution ofdi-tert-butyl-dicarbonate (478 mg) in tert-butanol (0.5 mL). Afterstirring overnight, the reaction mixture was diluted with ethyl acetate,washed with brine, dried (MgSO₄) and the solvent was removed in vacuo toyield (3R,5S)-3,5-dimethyl-piperazine-1-carboxylic acid tert-butyl ester(400 mg).

A mixture of (3R,5S)-3,5-dimethyl-piperazine-1-carboxylic acidtert-butyl ester (1.5 g), 2-bromoethyl methyl ether (1.32 mL) andpotassium carbonate (1.06 g) was heated to 120° C. in DMF (15 mL) for 2days. The reaction mixture was cooled, diluted with ethyl acetate,washed with brine, dried (MgSO₄) and the solvent was removed in vacuo toliberate(3R,5S)-4-(2-methoxy-ethyl)-3,5-dimethyl-piperazine-1-carboxylic acidtert-butyl ester (1.4 g) after column chromatography.

Removal of the BOC group with HCl yielded the desired compound, whichwas isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 1.01 (6H, d), 1.9 (2H, m), 2.61 (4H, m), 2.82(2H, t), 3.27 (3H, s), 3.37 (2H, t), 3.71 (2H, s), 3.85 (4H, m), 4.02(4H, m), 7.3 (1H, s), 7.43 (1H, t), 7.51 (1H, d), 8.21 (1H, d), 8.95(1H, s), 10.10 (1H, m); MS (ESI⁺) 522.35 (MH⁺).

1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylicacid (2-methoxy-ethyl)-methyl-amide (99)

Prepared via1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylicacid (2-methoxy-ethyl)-methyl-amide, prepared frompiperidine-4-carboxylic acid (2-methoxy-ethyl)-methyl-amide. The aminewas prepared as described for compound 100.

¹H NMR (400 MHz, CDCl₃): 1.71 (2H, m), 1.98 (2H, t), 2.18 (2H, m),2.46-2.70 (1H, m); 2.99+3.12 (3H, s, 2× rotamers), 3.08 (2H, m); 3.34(3H, s), 3.42-3.62 (4H, m), 3.86 (2H, s), 3.95 (4H, m), 4.10 (4H, m),7.36 (1H, s), 7.50 (1H, t), 7.58 (1H, d), 8.28 (1H, d), 9.01 (1H, s),10.07 (1H, br s); MS (ESI⁺) 550 (MH⁺).

1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylicacid dimethylamide (100)

Prepared via1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidine-4-carboxylicacid dimethylamide, prepared from piperidine-4-carboxylic aciddimethylamide.

Amine preparation: to a stirring solution of BOC-isonipecotic acid (400mg) in DMF (4 mL) was added 1,1′-carbonyldiimidazole (560 mg). Thereaction mixture was stirred overnight and then dimethylaminehydrochloride (280 mg) and triethylamine (0.48 mL) were added. After 5 hthe reaction mixture was diluted with ethyl acetate, washed with water,dried (MgSO₄) and the solvent was removed in vacuo to yield4-dimethylcarbamoyl-piperidine-1-carboxylic acid tert-butyl ester.Removal of the BOC group with HCl yielded the desired compound, whichwas isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 1.72 (2H, m), 1.98 (2H, m), 2.20 (2H, t), 2.55(1H, m), 2.97 (3H, s), 3.00-3.10 (5H, m), 3.86 (2H, s), 3.94 (4H, m),4.10 (4H, m), 7.36 (1H, s), 7.50 (1H, t, J=7.7 Hz), 7.60 (1H, d, J=8.2Hz), 8.28 (1H, d, J=7.3 Hz); 9.02 (1H, s); 10.15 (1H, br s); MS (ESI⁺)506 (MH⁺).

2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-3-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine(101)

Prepared via2-Chloro-4-morpholin-4-yl-6-(4-pyridin-3-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared from 1-pyridin-3-ylmethyl-piperazine. The amine was prepared asdescribed for compound 95.

¹H NMR (400 MHz, CDCl₃): 2.50-2.65 (m, 8H), 3.55 (s, 2H), 3.85 (s, 2H),3.90-3.93 (m, 4H), 4.07-4.10 (m, 4H), 7.24 (m, 1H), 7.36 (s, 1H), 7.49(t, 1H), 7.57 (d, 1H), 7.66 (d, 1H), 8.27 (d, 1H,), 8.50 (d, 1H), 8.54(s, 1H), 9.0 (s, 1H), 10.1 (sbr, H, NH); MS (ESI⁺) 527.25 (MH⁺).

1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylicacid methylamide (102)

Prepared via1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidine-4-carboxylicacid methylamide, prepared from piperidine-4-carboxylic acidmethylamide. The amine was prepared as described for compound 100.

¹H NMR (400 MHz, d₆-DMSO): 1.58-1.70 (4H, m), 2.00-2.15 (3H, m), 2.57(3H, d, J=4.5 Hz), 2.94 (2H, m), 3.84 (6H, m), 4.00 (4H, m), 7.46 (2H,t), 7.65 (2H, d), 8.20 (1H, d), 8.87 (1H, s); 13.14 (1H, s); MS (ESI⁺)492 (MH⁺).

2-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-N-methyl-isobutyramide(103)

Prepared via2-[4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazin-1-yl]-N-methyl-isobutyramide,prepared from N-methyl-2-piperazin-1-yl-isobutyramide.

Amine preparation: to a mixture of DCM (10 mL), aqueous sodiumbicarbonate (2M, 10 mL), and aqueous sodium carbonate (2M, 10 mL) wasadded methylamine hydrochloride (300 mg) at 0° C. To this was added2-bromoisobutyryl bromide (0.50 mL) with vigorous stirring. After 2 hagitation, standard work up yielded 2-bromo-2,N-dimethyl-propionamide(548 mg) as an off-white solid.

A mixture of 2-bromo-2,N-dimethyl-propionamide (312 mg),1-BOC-piperazine (323 mg) and silver oxide (800 mg) was stirred intoluene (5 mL) at reflux. After 24 h the reaction mixture was cooled,filtered through celite, diluted with chloroform, washed with brine,dried (MgSO₄) and the solvent was removed in vacuo to yield4-(1-methyl-1-methylcarbamoyl-ethyl)-piperazine-1-carboxylic acidtert-butyl ester (461 mg). Removal of the BOC group with HCl yielded thedesired compound, which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 10.1 (1H, br s), 9.02 (1H, s), 8.30 (1H, d),7.60 (1H, d), 7.51-7.49 (1H, m), 7.39 (1H, s), 7.20 (1H, q), 4.12-4.09(4H, m), 3.95-3.90 (4H, m), 3.87 (2H, s), 2.80 (3H, d), 2.65-2.50 (8H,m), 1.21 (6H, s); MS (ESI⁺) 535 (MH⁺).

2-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-2-methyl-1-pyrrolidin-1-yl-propan-1-one(104)

Prepared via2-[4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazin-1-yl]-2-methyl-1-pyrrolidin-1-yl-propan-1-one,prepared from 2-methyl-2-piperazin-1-yl-1-pyrrolidin-1-yl-propan-1-one.

Amine preparation: to pyrrolidine (390 uL) in dry THF (4 mL) at 0° C.was added nBuLi (1.86 ml) The reaction mixture was stirred for 5 minutesand then 4-(1-ethoxycarbonyl-1-methyl-ethyl)-piperazine-1-carboxylicacid tert-butyl ester (700 mg) in THF (5 mL) was added. The reactionmixture was stirred at room temperature for 24 h and then quenched withaqueous ammonium chloride, extracted into DCM, washed with water, dried(MgSO₄) and the solvent removed in vacuo to yield4-(1,1-dimethyl-2-oxo-2-pyrrolidin-1-yl-ethyl)-piperazine-1-carboxylicacid tert-butyl ester (745 mg). Removal of the BOC group with HClyielded the desired compound, which was isolated as the hydrochloridesalt.

¹H NMR (400 MHz, CDCl₃): 1.24 (6H, s), 1.79-1.92 (4H, m), 2.52-2.66 (8H,br), 3.49 (2H, t), 3.82 (2H, s), 3.92 (4H, t), 4.00 (2H, t), 4.05 (4H,t), 7.35 (1H, s), 7.51 (1H, t), 7.59 (1H, d), 8.29 (1H, d), 9.03 (1H,s), 10.10 (1H, br); MS (ESI⁺) 575 (MH⁺).

2-(1H-Indazol-4-yl)-6-[4-(1-methyl-1H-imidazol-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(105)

Prepared via2-chloro-6-[4-(1-methyl-1H-imidazol-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-(1-methyl-1H-imidazol-2-ylmethyl)-piperazine. The aminewas prepared as described for compound 95.

¹H NMR (400 MHz, CDCl₃): 2.50-2.65 (8H, br), 3.63 (2H, s), 3.71 (3H, s),3.85 (2H, s), 3.92 (4H, t), 4.05 (4H, t), 6.85 (1H, s), 6.92 (1H, s),7.38 (1H, s), 7.51 (1H, t), 7.60 (1H, d), 8.29 (1H, d), 9.02 (1H, s); MS(ESI⁺) 530 (MH⁺).

2-(1H-Indazol-4-yl)-6-[4-(5-methyl-isoxazol-3-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(106)

Prepared via2-Chloro-6-[4-(5-methyl-isoxazol-3-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-(5-methyl-isoxazol-3-ylmethyl)-piperazine.

Amine preparation: To a suspension of lithium aluminium hydride (1.04 g)in THF (10 mL) was added methyl 5-methylisoxazole-3-carboxylate (1.00 g)as a solution in THF (10 mL). After 24 h the reaction mixture wasquenched with aqueous ammonium chloride, extracted into ethyl acetate,dried (MgSO₄) and the solvent was removed in vacuo to yield(5-methyl-isoxazol-3-yl)-MeOH (579 mg).

To a solution of (5-methyl-isoxazol-3-yl)-MeOH (570 mg) in DCM (15 mL)was added triethylamine (0.98 mL) followed by methanesulfonyl chloride(0.51 mL). After 1 h the reaction mixture was diluted with DCM, washedwith water, dried (MgSO₄) and the solvent was removed in vacuo to yieldmethanesulfonic acid 5-methyl-isoxazol-3-ylmethyl ester (887 mg).

To a solution of N—BOC-piperazine (300 mg) in MeCN (5 mL) was addedpotassium carbonate (289 mg) followed by methanesulfonic acid5-methyl-isoxazol-3-ylmethyl ester (369 mg). The reaction mixture washeated to reflux for 24 h. After cooling the reaction mixture wasdiluted with water extracted into ethyl acetate, dried (MgSO₄) and thesolvent was removed in vacuo to yield4-(5-methyl-isoxazol-3-ylmethyl)-piperazine-1-carboxylic acid tert-butylester (404 mg).

Removal of the BOC group with HCl yielded the desired compound, whichwas isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 2.41 (s, 3H), 2.59 (br s, 8H), 3.59 (s, 2H),3.85 (s, 2H), 3.92 (t, 4H), 4.09 (t, 4H), 5.99 (s, 1H), 7.37 (s, 1H),7.50 (t, 1H), 7.58 (d, 1H), 8.28 (d, 1H), 9.02 (s, 1H), 10.15 (br s,1H); MS (ESI⁺) 531 (MH⁺).

1-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-2-methyl-propan-2-ol(107)

Prepared via1-[4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazin-1-yl]-2-methyl-propan-2-ol,prepared from 2-methyl-1-piperazin-1-yl-propan-2-ol.

Amine preparation: a mixture of 1-benzylpiperazine (5 g), ethylbromoacetate (3.15 mL) and potassium carbonate (4.31 g) was stirred atroom temperature in MeCN (50 mL). After stirring overnight the reactionmixture was diluted with chloroform, washed with brine, dried (MgSO₄)and the solvent removed in vacuo to yield4-ethoxycarbonylmethyl-piperazine-1-carboxylic acid tert-butyl ester(5.86 g)

To 4-ethoxycarbonylmethyl-piperazine-1-carboxylic acid tert-butyl ester(1.0 g) in dry THF (10 mL) at 0° C. was added methyl magnesium bromide(3.0M solution in diethyl ether, 8.6 mL). The reaction mixture was thenheated to reflux for 24 h. The reaction mixture was poured intoice/water/brine and then extracted into ethyl acetate, dried (MgSO₄) andthe solvent removed in vacuo to yield4-(2-hydroxy-2-methyl-propyl)-piperazine-1-carboxylic acid tert-butylester. Transfer hydrogenation with ammonium formate and 10% palladium oncarbon in MeOH afforded the desired compound.

¹H NMR (400 MHz, CDCl₃): 1.17 (6H, s), 2.36 (2H, s), 2.62 (4H, m), 2.73(4H, m), 3.86 (2H, s), 3.92 (4H, m), 4.1 (4H, m), 7.38 (1H, s), 7.50(1H, t), 7.58 (1H, d), 8.28 (1H, d), 9.02 (1H, s); MS (ESI⁺) 508 (MH⁺).

Cyclopropylmethyl-{1-[2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-amine(108)

Prepared via[1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-cyclopropylmethyl-(2-methoxy-ethyl)-amine,prepared from cyclopropylmethyl-(2-methoxy-ethyl)-piperidin-4-yl-amine.

Amine preparation: 1-BOC-4-piperidone (500 mg) and 2-methoxyethylamine(218 μL) were stirred in MeOH at room temperature. After 16 h, sodiumborohydride was added (190 mg) carefully. After a further 3 h, thereaction mixture was diluted with DCM, washed with water, dried (MgSO₄)and the solvent removed in vacuo to yield4-(2-methoxy-ethylamino)-piperidine-1-carboxylic acid tert-butyl ester(560 mg).

A mixture of 4-(2-methoxy-ethylamino)-piperidine-1-carboxylic acidtert-butyl ester (525 mg), cyclopropylmethyl bromide (218 μL) andpotassium carbonate (340 mg) was heated to reflux in MeCN for 16 h.After cooling the reaction mixture was diluted with chloroform, washedwith brine, dried (MgSO₄) and the solvent removed in vacuo to yield4-[cyclopropylmethyl-(2-methoxy-ethyl)-amino]-piperidine-1-carboxylicacid tert-butyl ester (475 mg). Removal of the BOC group with HClyielded the desired compound, which was isolated as the hydrochloridesalt.

¹H NMR (400 MHz, CDCl₃): −0.01-0.01 (2H, m), 0.40-0.48 (2H, m),1.45-1.60 (3H, m), 1.62-1.70 (2H, m), 1.97-2.04 (2H, m), 2.33 (2H, d),2.52-2.61 (1H, m), 2.67 (2H, t), 2.92-3.00 (2H, m), 3.25 (3H, s), 3.34(2H, t), 3.71 (2H, s), 3.82 (4H, t), 4.00 (4H, t), 7.22 (1H, s), 7.49(1H, t), 7.48 (1H, d), 8.28 (1H, d), 8.90 (1H, s), 10.00 (1H, br); MS(ESI⁺) 562 (MH+).

6-[4-(1-Ethyl-1-methoxymethyl-propyl)-piperazin-1-ylmethyl]-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(109)

Prepared via2-Chloro-6-[4-(1-ethyl-1-methoxymethyl-propyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-(1-ethyl-1-methoxymethyl-propyl)-piperazine.

To a solution of 1-(4-benzyl-piperazin-1-yl)-2-methoxy-ethanone (2.60 g)in dry THF (30 mL) at −10° C. was added titanium isopropoxide (3.22 mL),followed by ethyl magnesium bromide (1.0M solution in THF, 22.05 mL).The reaction mixture was heated to reflux for 2 days. After cooling thereaction mixture was cooled, diluted with chloroform, washed with brine,dried (MgSO₄) and the solvent removed in vacuo. The residue was purifiedusing flash chromatography to yield1-benzyl-4-(1-methoxymethyl-cyclopropyl)-piperazine (452 mg) and1-benzyl-4-(1-ethyl-1-methoxymethyl-propyl)-piperazine (248 mg).Transfer hydrogenation conditions using ammonium formate and 10% Pd/C inMeOH afforded the desired compound.

¹H NMR (400 MHz, CDCl₃) 0.85 (6H, t), 1.58-1.33 (4H, m), 2.55-2.50 (4H,m), 2.75-2.70 (4H, m), 3.30 (5H, s), 3.85 (2H, s), 3.95-3.91 (4H, m),4.12-4.09 (4H, m), 7.39 (1H, s), 7.51-7.49 (1H, m), 7.60 (1H, d), 8.30(1H, d), 9.02 (1H, s), 10.2 (1H, br s); MS (ESI⁺) 550 (MH⁺).

2-(1H-Indazol-4-yl)-6-[4-(1-methoxymethyl-cyclopropyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine(110)

Prepared via2-Chloro-6-[4-(1-methoxymethyl-cyclopropyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-(1-methoxymethyl-cyclopropyl)-piperazine. Amine wasprepared as outlined for compound 109.

¹H NMR (400 MHz, CDCl₃) 0.65-0.52 (4H, m), 2.55-2.50 (4H, m), 2.85-2.80(4H, m), 3.32 (3H, s), 3.40 (2H, s), 3.85 (2H, s), 3.95-3.91 (4H, m),4.12-4.09 (4H, m), 7.39 (1H, s), 7.51-7.49 (1H, m), 7.60 (1H, d), 8.30(1H, d), 9.02 (1H, s), 10.2 (1H, br s); MS (ESI⁺) 520 (MH⁺).

111{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-(2,2,2-trifluoro-ethyl)-amine

Via[1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-(2-methoxy-ethyl)-(2,2,2-trifluoro-ethyl)-amine,prepared from(2-methoxy-ethyl)-piperidin-4-yl-(2,2,2-trifluoro-ethyl)-amine.

Amine preparation: 1-BOC-4-piperidinone (2.00 g) and 2-methoxyethylamine(872 μL) were stirred together in MeOH (20 mL) at room temperatureovernight. Sodium borohydride (760 mg) was then added portionwise andthe reaction mixture was allowed to stir further at ambient temperature.After 16 h, the solvent was removed in vacuo, the residue was dilutedwith DCM, washed with brine, dried (MgSO₄) and the solvent removed invacuo. The residue was purified using flash chromatography to yield4-(2-methoxy-ethylamino)-piperidine-1-carboxylic acid tert-butyl esteras a colourless oil (1.69 g).

To a solution of 4-(2-methoxy-ethylamino)-piperidine-1-carboxylic acidtert-butyl ester (500 mg) in DCM (5 mL) and triethylamine (540 μL) wasadded trifluoroacetic anhydride (548 μL). The reaction mixture wasstirred at room temperature overnight, diluted with DCM, washed withbrine, dried (MgSO₄) and the solvent removed in vacuo. The residue waspurified using flash chromatography to yield4-[(2-methoxy-ethyl)-(2,2,2-trifluoro-acetyl)-amino]-piperidine-1-carboxylicacid tert-butyl ester as an oil (685 mg).

To a solution of4-[(2-methoxy-ethyl)-(2,2,2-trifluoro-acetyl)-amino]-piperidine-1-carboxylicacid tert-butyl ester (685 mg) in dry THF (7 mL) was added borane-methylsulfide complex (405 uL) at 0° C. under inert atmosphere. The reactionmixture was refluxed for 3 h, and then stirred at room temperatureovernight, quenched with MeOH, diluted with DCM, washed with brine,dried (MgSO₄) and the solvent removed in vacuo. The residue was purifiedusing flash chromatography to yield4-[(2-methoxy-ethyl)-(2,2,2-trifluoro-ethyl)-amino]-piperidine-1-carboxylicacid tert-butyl ester as an oil (635 mg). Treatment of this compoundwith HCl in DCM/MeOH furnished the desired amine, isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.55-1.64 (2H, m), 1.76-1.82 (2H, m), 2.12-2.18(2H, m), 2.58-2.62 (1H, m), 2.86 (2H, t, J=6.3 Hz), 3.03-3.08 (2H, m),3.20 (2H, q, J=9.4 Hz), 3.33 (3H, s), 3.45 (2H, t, J=6.4 Hz), 3.84 (2H,s), 4.00 (4H, t, J=5.1 Hz), 7.22 (1H, s), 7.49 (1H, t, J=7.2 Hz), 7.48(1H, d, J=8.3 Hz), 8.28 (1H, d, J=7.1 Hz), 8.90 (1H, s), 10.00 (1H, br);MS (ESI⁺) 590 (MH⁺).

1212-(1H-Indazol-4-yl)-6-[4-(2-methoxy-ethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-Chloro-6-[4-(2-methoxy-ethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-(2-methoxy-ethyl)-piperazine.

¹H NMR (400 MHz, CDCl₃) 2.55-2.70 (m, 10H, 5×CH₂), 3.35 (s, 2H, CH₂),3.50-3.53 (m, 2H, CH₂), 3.85 (s, 2H, CH₂), 3.90-3.93 (m, 4H, 2×CH₂),4.07-4.11 m, 4H, 2×CH₂), 7.37 (s, H, ArH), 7.49 (t, H, ArH, J=7.76 Hz),7.57 (d, H, ArH, J=8.3 Hz), 8.27 (d, H, ArH, J=6.71 Hz), 9.0 (s, H,ArH), 10.15 (br s, H, NH). MS (ESI⁺) 494.18 (MH⁺).

1222-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-isobutyramide

Via2-[4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazin-1-yl]-isobutyramide,prepared from 2-piperazin-1-yl-isobutyramide.

Amine preparation: a mixture of 1-BOC-piperazine (1.063 g), DCM/MeOH (20mL) and 2.0M HCl in ether (3.14 mL) was stirred at 0° C. After 1 hr thesolvent was removed in vacuo to yield a white solid. This was dissolvedin water and sodium cyanide (280 mg) was added. To this was added asolution of acetone (420 μL) in water 2 mL. After stirring for 48 hrsthe reaction mixture was dissolved in ethyl acetate, washed with brine,dried (MgSO₄) and the solvent removed in vacuo to yield4-(cyano-dimethyl-methyl)-piperazine-1-carboxylic acid tert-butyl ester(1.11 g).

To a cooled (ice/water bath) solution of4-(cyano-dimethyl-methyl)-piperazine-1-carboxylic acid tert-butyl ester(102.9 mg) in dry DMSO was added K₂CO₃ (9.8 mg) followed by 28% hydrogenperoxide solution (200 μL) dropwise. The resultant was heated at 40° C.overnight. Water was added and the white solid collected, washed withwater and air dried to give4-(1-carbamoyl-1-methyl-ethyl)-piperazine-1-carboxylic acid tert-butylester (59.6 mg). (See Tetrahedron 2002, 58, 3217). Treatment of thiscompound with HCl in DCM/MeOH yielded the desired amine, which wasisolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.24 (s, 6H, 2×CH₂), 2.55-2.65 (m, 8H, 4×CH₂),3.85 (s, 2H, CH₂), 3.90-3.92 (m, 4H, 2×CH₂), 4.07-4.09 (m, 4H, 2×CH₂),5.35 (m, H, NH), 7.09 (m, H, NH), 7.37 (s, H, ArH), 7.48 (t. H, ArH,J=7.72 Hz), 7.57 (d, H, ArH, J=8.22 Hz), 8.26 (d, H, ArH, J=7.14 Hz),9.0 (s, H, ArH, 10.4 (br s, H, NH). MS (ESI⁺) 521.27 (MH⁺).

123{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-methanol

Via[1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-methanol,prepared from piperidin-4-yl-methanol.

Amine preparation: To a solution of piperidine-4-carboxylic acid ethylester (5.0 g) stirring in dry MeCN (70 mL) was added triethylamine (5.3mL), followed by di-tertiary-butyl dicarbonate (7.64 g). The reactionmixture was stirred at room temperature for 24 h, then diluted withwater (150 mL) extracted into ethyl acetate, dried (MgSO₄) and thesolvent removed in vacuo. The residue was purified using flashchromatography to give 1-BOC-piperidine-4-ethyl ester (6.8 g) as acolourless oil. To a suspension of lithium aluminium hydride powder (300mg) stirring in dry THF (10 mL) under a nitrogen atmosphere was added1-BOC-piperidine-4-ethyl ester dissolved in dry THF (5 mL). The reactionmixture was stirred at room temperature. After 2 h the reaction mixturewas quenched with saturated ammonium chloride solution (10 mL), filteredthrough a celite bed and the solvent removed in vacuo to yield4-hydroxymethyl-piperidine-1-carboxylic acid tert-butyl ester (0.793 g)as a pale yellow solid. Treatment with TFA yielded the desired amine,which was isolated as the TFA salt.

¹H NMR (400 MHz, d₆-DMSO) 1.18 (m, 2H), 1.37 (m, 1H), 1.64 (m, 2H), 2.04(t, 2H), 2.92 (m, 2H), 3.27 (m, 2H), 3.82 (m, 4H & CH₂), 3.99 (m, 4H),4.39 (t, 1H), 7.45 (t, 2H), 7.65 (d, 1H, J=8.2 Hz), 8.21 (d, 1H, J=7.3Hz), 8.87 (s, 1H), 13.16 (br s, 1H); MS (ESI⁺) 465.17 (MH⁺).

1242-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-4-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine

Via2-Chloro-4-morpholin-4-yl-6-(4-pyridin-4-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared from 1-pyridin-4-ylmethyl-piperazine.

Amine preparation: a mixture of 1-BOC-piperazine (2 g), 4-pyridinecarboxaldehyde (1.26 g), sodium triacetoxyborohydride (2.96 g) andacetic acid (0.6 mL) was stirred together in dry 1,2-dichloroethane (15mL) at room temperature. After 4 h the reaction was quenched withsaturated NaHCO₃ solution and extracted with DCM, dried (Na₂SO₄),filtered and concentrated in vacuo to give4-pyridin-4-ylmethyl-piperazine-1-carboxylic acid tert-butyl ester (3g). Treatment of this compound with HCl in DCM/MeOH yielded the desiredamine, which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 2.40-2.60 (m, 8H, 4×CH₂), 3.45 (s, 2H, CH₂),3.78 (s, 2H, CH₂), 3.82-3.84 (m, 4H, 2×CH₂), 3.99-4.01 (m, 4H, 2×CH₂),7.18-7.19 (m, 2H, 2×ArH), 7.30 (s, H, ArH), 7.39 (t, H, ArH, J=7.7 Hz),7.47 (d, H, ArH, J=8.55 Hz), 8.18 (d, H, ArH, J=7.17 Hz), 8.46 (d, H,ArH, J=5.34 Hz), 8.93 (s, H, ArH), 10.25 (br s, H, NH); MS (ESI⁺) 527.29(MH⁺).

1252-(1H-Indazol-4-yl)-6-[4-(6-methyl-pyridin-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-chloro-6-[4-(6-methyl-pyridin-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-(6-methyl-pyridin-2-ylmethyl)-piperazine. The amine,1-(6-methyl-pyridin-2-ylmethyl)-piperazine, was prepared using6-methyl-2-pyridinecarboxaldehyde, analogous to 124.

¹H NMR (400 MHz, CDCl₃) 2.54 (s, 3H, CH₃), 2.60-2.70 (m, 8H, 4×CH₂),3.67 (s, 2H, CH₂), 3.86 (s, 2H, CH₂), 3.90-3.93 (m, 4H, 2×CH₂),4.07-4.10 (m, 4H, 2×CH₂), 7.00 (d, H, ArH, J=7.6 Hz), 7.22-7.25 (m, H;ArH part under CDCl₃), 7.36 (s, H, ArH), 7.47-7.57 (m, 3H, 3×ArH), 8.27(d, H, ArH, J=7.2 Hz), 9.00 (s, H, ArH), 10.2 (br s, H. NH); MS (ESI⁺)541.24 (MH⁺).

1262-(1H-Indazol-4-yl)-6-[4-(4-methyl-thiazol-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-chloro-6-[4-(4-methyl-thiazol-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-(4-methyl-thiazol-2-ylmethyl)-piperazine. The amine,1-(4-methyl-thiazol-2-ylmethyl)-piperazine, was prepared using4-methylthiazole-2-carboxaldehyde, analogous to 125.

¹H NMR (400 MHz, CDCl₃) 2.43 (s, 3H), (br s, 8H), 3.85 (m, 4H), 3.92 (m,4H), 4.09 (m, 4H), 6.83 (s, 1H), 7.38 (s, 1H), 7.51 (m, 1H), 7.60 (d,J=8.3 Hz, 1H), 8.28 (d, J=6.8 Hz, 1H), 9.02 (s, 1H); MS (ESI⁺) 547(MH⁺).

127{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-pyridin-2-yl-amine

Via[1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-pyridin-2-yl-amine,prepared from piperidin-4-yl-pyridin-2-yl-amine.

Amine preparation: A mixture of 1-BOC-4-piperidone (496 mg),2-aminopyridine (234 mg), sodium triacetoxyborohydride (580 mg) andacetic acid (0.14 mL) was stirred together in dry 1,2-dichloroethane (10mL) at room temperature. After 24 h the reaction was quenched withsaturated NaHCO₃ solution and extracted with DCM, dried (Na₂SO₄),filtered and concentrated in vacuo to give4-(pyridin-2-ylamino)-piperidine-1-carboxylic acid tert-butyl ester.(135 mg) after flash chromatography. Treatment of this compound with HClin DCM/MeOH yielded the desired amine, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.65-1.55 (2H, m), 2.10-2.08 (2H, m), 2.40-2.30(2H, m), 3.01 (2H, d), 3.73-3.67 (1H, m), 3.85 (2H, s), 3.95-3.91 (4H,m), 4.12-4.09 (4H, m), 4.40 (1H, br d), 6.39 (1H, d), 6.56-6.521 (1H,m), 7.48-7.40 (2H, m), 7.51-7.49 (1H, m), 7.60 (1H, d), 8.09 (1H, d),8.30 (1H, d), 9.02 (1H, s), 10.2 (1H, br s); MS (ESI⁺) 527 (MH⁺).

128N-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-2-methoxy-N-methyl-acetamide.

ViaN-[1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-2-methoxy-N-methyl-acetamide,prepared from 2-methoxy-N-methyl-N-piperidin-4-yl-acetamide.

Amine preparation: To a solution of 1-BOC-4-piperidone (1.0 g) in MeOH(10 mL) was added a solution of freshly prepared methylamine in MeOH(1.0 mL). The reaction mixture was stirred for 1 h and then sodiumcyanoborohydride (0.315 g) was added. After stirring for 24 h thereaction mixture was then diluted with DCM, washed with sodiumbicarbonate solution, dried (MgSO₄) and the solvent removed in vacuo.The residue was purified by flash chromatography to yield4-methylamino-piperidine-1-carboxylic acid tert-butyl ester (0.85 g).

To a solution of 4-methylamino-piperidine-1-carboxylic acid tert-butylester (0.42 g) in DCM (10 mL) was added triethylamine (0.30 mL) followedby methoxyacetyl chloride (0.20 mL). After stirring for 3 h the reactionmixture was then diluted with DCM, washed with sodium bicarbonatesolution, dried (MgSO₄) and the solvent removed in vacuo. The residuewas purified by flash chromatography to yield4-[(2-methoxy-acetyl)-methyl-amino]-piperidine-1-carboxylic acidtert-butyl ester (0.293 g). Treatment of this compound with HCl inDCM/MeOH yielded the desired amine, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.66 (2H, m), 1.80+1.97 (2H, m, 2 rotamers),2.36 (2H, m), 2.90 (3H, s), 3.08 (2H, m), 3.43 (3H, s), 3.65+4.50 (1H,m, 2 rotamers), 3.86 (2H, s), 3.94 (4H, m), 4.10 (4H, m), 7.40 (1H, s),7.51 (1H, t, J=7.7 Hz), 7.54 (1H, d, J=8.2 Hz), 8.28 (1H, d, J=7.2 Hz),9.02 (1H, s), 10.18 (1H, br s); MS (ESI⁺) 536 (MH⁺).

129N-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-N-methyl-methanesulfonamide

ViaN-[1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-N-methyl-methanesulfonamide,prepared from N-methyl-N-piperidin-4-yl-methanesulfonamide. The amine,N-methyl-N-piperidin-4-yl-methanesulfonamide, was prepared by treatmentof 4-methylamino-piperidine-1-carboxylic acid tert-butyl ester withmethanesulfonyl chloride, analogous to 128.

¹H NMR (400 MHz, CDCl₃) 1.72 (2H, m), 1.90 (2H, m), 2.23 (2H, t, J=11.0Hz), 2.85 (6H, s), 3.08 (2H, br d, J=11.4 Hz), 3.80 (1H, m), 3.86 (2H,s), 3.94 (4H, m), 4.10 (4H, m), 7.39 (1H, s), 7.52 (1H, t, J=7.7 Hz),7.60 (1H, d, J=8.0 Hz), 8.29 (1H, d, J=7.1 Hz), 9.03 (1H, s), 10.15 (1H,br s); MS (ESI⁺) 542 (MH⁺).

130{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]piperidin-4-yl}-(3-methoxy-propyl)-methyl-amine

Via[1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-(3-methoxy-propyl)-methyl-amine,prepared from (3-methoxy-propyl)-methyl-piperidin-4-yl-amine. The amine,(3-methoxy-propyl)-methyl-piperidin-4-yl-amine, was prepared from4-methylamino-piperidine-1-carboxylic acid tert-butyl ester andtoluene-4-sulfonic acid 3-methoxy-propyl ester (produced from3-methoxy-1-propanol using standard conditions) in a similar manner to128.

¹H NMR (400 MHz, CDCl₃) 1.55-1.85 (6H, 3×CH₂), 2.11 (m, 2H, CH₂), 2.31(s, 3H, CH₃), 2.44 (m, H, CH), 2.57 (m, 2H, CH₂), 3.05 (m, 2H, CH₂),3.33 (s, 3H, CH₃), 3.42 (m, 2H, CH₂), 3.83 (s, 2H, CH₂), 3.91-3.93 (m,4H, 2×CH₂), 4.08-4.10 (m, 4H, 2×CH₂), 7.35 (s, H, ArH), 7.50 (t, H, ArH,J=7.77 Hz), 7.58 (d, H, ArH, J=8.27 Hz), 8.26 (d, H, ArH, J=6.77 Hz),9.00 (s, H, ArH), 10.23 (s br, H, NH); MS (ESI⁺) 536.46 (MH⁺).

1316-((3S,5R)-3,5-Dimethyl-4-pyridin-2-ylmethyl-piperazin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[312-d]pyrimidine

Via2-Chloro-6-((3S,5R)-3,5-dimethyl-4-pyridin-2-ylmethyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from (2S,6R)-2,6-dimethyl-1-pyridin-2-ylmethyl-piperazine.Amine preparation: (3R,5S)-3,5-dimethyl-piperazine-1-carboxylic acidtert-butyl ester (845 mg), 2-(bromomethyl)-pyridine hydrobromide (1 g)and potassium carbonate (1.15 g) was heated to reflux in MeCN (10 mL).After heating for 24 h the reaction mixture was diluted with DCM, washedwith sodium bicarbonate solution, dried (MgSO₄) and the solvent removedin vacuo. The residue was purified by flash chromatography to yield(3S,5R)-3,5-dimethyl-4-pyridin-2-ylmethyl-piperazine-1-carboxylic acidtert-butyl ester (867 mg). Treatment of this compound with HCl inDCM/MeOH yielded the desired amine, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.00 (d, J=6.0 Hz, 6H), 1.54 (s, 2H), 2.05 (m,2H), 2.84 (m, 4H), 3.81 (s, 2H), 3.92 (m, 4H), 4.10 (m, 4H), 7.11 (m,1H), 7.38 (s, 1H), 7.51 (m, 1H), 7.61 (m, 3H), 8.29 (d, J=7.4 Hz, 1H),8.51 (d, J=4.5 Hz, 1H), 9.03 (s, 1H); MS (ESI⁺) 555 (MH⁺).

1322-(1H-Indazol-4-yl)-6-(4-methoxymethyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-Chloro-6-(4-methoxymethyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 4-methoxymethyl-piperidine. Amine preparation: To asolution of 4-hydroxymethyl-piperidine-1-carboxylic acid tert-butylester (212 mg) stirring in dry THF (10 mL) under a nitrogen atmospherewas added sodium hydride (60% in paraffin oil; 45 mg) followed after 20min by methyl iodide and the reaction mixture was warmed to 70° C. After4 h the mixture was cooled and quenched with water (10 mL) extractedinto ethyl acetate, and dried (MgSO₄). The residue was purified usingflash chromatography to yield 4-methoxy-piperidine-1-carboxylic acidtert-butyl ester (158 mg) as a colourless oil. Treatment with TFAyielded the desired amine which was isolated as the TFA salt.

¹H NMR (400 MHz, CDCl₃) 1.23 (m, 2H), 1.54 (m, 1H), 1.63 (m, 2H), 2.05(m, 2H), 2.91 (m, 2H), 3.18 (d, 2H, J=6.2 Hz), 3.22 (s, 3H), 3.82 (m,4H, +CH₂), 3.99 (m, 4H), 7.45 (t, 2H, J=7.0 Hz), 7.65 (d, 1H, J=8.2 Hz),8.21 (d, 1H, J=7.1 Hz), 8.87 (s, 1H), 13.16 (br s, 1H); MS (ESI⁺) 479.2(MH⁺).

133{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-thiazol-2-ylmethyl-amine

Via[1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-(2-methoxy-ethyl)-thiazol-2-ylmethyl-amine,prepared from (2-methoxy-ethyl)-piperidin-4-yl-thiazol-2-ylmethyl-amine.

Amine preparation: 4-(2-Methoxy-ethylamino)-piperidine-1-carboxylic acidtert-butyl ester (465 mg) and 2-thiazolecarboxaldehyde (190 ul) werestirred in dry 1,2-dichloroethane (5 mL) for 1 h. Next were added aceticacid (1 eq.) and sodium triacetoxyborohydride (458 mg). The reactionmixture was stirred at room temperature overnight, diluted with DCM,washed with brine, dried (MgSO₄) and the solvent removed in vacuo. Theresidue was purified using flash chromatography to yield4-[(2-methoxy-ethyl)-thiazol-2-ylmethyl-amino]-piperidine-1-carboxylicacid tert-butyl ester (574 mg). Treatment of this compound with HCl inDCM/MeOH and basic wash with sodium hydrogen carbonate yielded thedesired amine.

¹H NMR (400 MHz, CDCl₃) 1.62-1.73 (2H, m), 1.81-1.88 (2H, m), 2.06-2.14(2H, m), 2.60-2.68 (1H, m), 2.88 (2H, t, J=6.4 Hz), 3.02-3.08 (2H, m),3.30 (3H, s), 3.49 (2H, t, J=6.4 Hz), 3.82 (2H, s), 3.92-3.96 (4H, m),4.10 (2H, s), 4.10-4.14 (4H, m), 7.22 (1H, d, J=3.2), 7.35 (1H, s), 7.51(1H, t, J=8.0 Hz), 7.59 (1H, d, J=8.3), 7.72 (1H, d, J=3.2), 8.29 (1H,d, J=6.6 Hz), 9.03 (1H, s), 10.10 (1H, br); MS (ESI⁺) 605 (MH⁺).

1341-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-4-pyridin-2-ylmethyl-piperidin-4-ol

Via1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-4-pyridin-2-ylmethyl-piperidin-4-ol,prepared from 4-pyridin-2-ylmethyl-piperidin-4-ol.

Amine preparation: To a solution of 2-picoline (333 mg) in dry THF (5mL) at −78° C. was added nBuLi (2.5M solution in hexanes, 1.50 mL). Thereaction mixture was warmed to room temperature over 30 min and thencooled to −78° C. 1-BOC-4-piperidone (713 mg) was then added, and thereaction mixture was raised to 0° C. over 20 min. The reaction mixturewas stirred overnight and then quenched with water. The organics wereextracted into ethyl acetate, dried (MgSO₄), filtered and concentratedin vacuo to give crude product. The residue was purified using flashchromatography to yield4-hydroxy-4-pyridin-2-ylmethyl-piperidine-1-carboxylic acid tert-butylester (290 mg). Treatment of this compound with HCl in DCM/MeOH yieldedthe desired amine, which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.55-1.75 (4H, m), 2.55-2.61 (2H, m), 2.68-2.72(2H, m), 2.91 (2H, s), 3.88 (2H, s), 3.91-3.94 (4H, m), 4.05-4.10 (4H,m), 5.80 (1H, s), 7.05-7.11 (2H, m), 7.37 (1H, s), 7.46-7.51 (1H, s),7.55-7.62 (2H, m), 8.29 (1H, d), 8.50 (1H, d), 9.03 (1H, s), 10.10 (1H,br s); MS (ESI⁺) 542 (MH⁺).

135{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-isopropyl-(2-methoxy-ethyl)-amine

Via[1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-isopropyl-(2-methoxy-ethyl)-amine,prepared from isopropyl-(2-methoxy-ethyl)-piperidin-4-yl-amine.

Amine preparation: A mixture of4-(2-methoxy-ethylamino)-piperidine-1-carboxylic acid tert-butyl ester(see preparation of 121) (300 mg) and 2-bromopropane (1.20 mL) in MeCN(3 mL) with potassium carbonate (192 mg) were heated at 60° C. in asealed tube for 7 days. The reaction mixture was cooled down, dilutedwith DCM, washed with brine, dried (MgSO₄) and the solvent removed invacuo. The residue was purified using flash chromatography to yield4-[isopropyl-(2-methoxy-ethyl)-amino]-piperidine-1-carboxylic acidtert-butyl ester as an oil (131 mg). Treatment of this compound with HClin DCM/MeOH and basic wash with aqueous sodium bicarbonate yielded thedesired amine.

¹H NMR (400 MHz, CDCl₃) 1.03 (6H, d, J=6.6 Hz), 1.62-1.72 (4H, m),2.08-2.14 (2H, m), 2.52-2.60 (1H, m), 2.69 (2H, t, J=7.4 Hz), 3.03-3.12(4H, m), 3.33 (2H, t, J=7.3 Hz), 3.35 (3H, s), 3.82 (2H, s), 3.92 (4H,t, J=4.5 Hz), 4.05 (4H, t, J=4.5 Hz), 7.35 (1H, s), 7.51 (1H, t, J=8.0Hz), 7.59 (1H, d, J=8.3 Hz), 8.29 (1H, d, J=6.6 Hz), 9.03 (1H, s), 10.10(1H, br); MS (ESI⁺) 550 (MH⁺).

1362-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(pyridin-2-yloxy)-piperidin-1-ylmethyl]-thieno[3,2-d]pyrimidine

Via2-chloro-4-morpholin-4-yl-6-[4-(pyridin-2-yloxy)-piperidin-1-ylmethyl]-thieno[3,2-d]pyrimidine,prepared from 2-(piperidin-4-yloxy)-pyridine.

Amine preparation: A mixture of 1-benzyl-piperidin-4-ol (1 g),2-chloropyridine (0.5 mL), 18-crown-6 (72 mg) and KOH (290 mg) wasrefluxed in dry toluene. After 18 h, the reaction mixture was dilutedwith water, extracted with DCM, dried (Na₂SO₄), filtered andconcentrated in vacuo then purified by flash chromatography to give4-(pyridin-2-yloxy)-piperidine-1-carboxylic acid tert-butyl ester (789mg). Treatment of this compound with hydrogen, 10% palladium on carbonand ammonium formate in MeOH yielded the desired amine.

¹H NMR (400 MHz, CDCl₃): 1.88-1.90 (m, 2H, 2×CH), 2.04-2.08 (m, 2H,2×CH), 3.48-2.52 (m, 2H, 2×CH), 2.85-2.90 (m, 2H, 2×CH), 3.88 (s, 2H,CH₂), 3.91-3.93 (m, 4H, 2×CH₂), 4.08-4.11 (m, 4H, 2×CH₂), 5.10-5.18 (m,H, CH), 6.71 (d, H, ArH, J=8.34 Hz), 6.82 (t, H, ArH, J=6.11 Hz), 7.37(s, H, ArH), 7.47-7.58 (m, 3H, 3×ArH), 8.12 (d, H, ArH, J=5.01 Hz), 8.27(d, H, ArH, J=6.8 Hz), 9.01 (s, H, ArH), 10.09 (br s, H, NH); MS (ESI⁺)528.31 (MH⁺).

137N-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-N-(2-methoxy-ethyl)-methanesulfonamide

ViaN-[1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-N-(2-methoxy-ethyl)-methanesulfonamide,prepared from N-(2-methoxy-ethyl)-N-piperidin-4-yl-methanesulfonamide.

Amine preparation: To a solution of4-(2-methoxyethylamine)-piperidine-1-carboxylic acid tert-butyl ester(see preparation of 121) (0.50 g) in DCM (10 mL) was added triethylamine(0.30 mL) followed by methanesulfonyl chloride (0.16 mL). After stirringfor 4 h the reaction mixture was then diluted with DCM, washed withsodium bicarbonate solution, dried (MgSO₄) and the solvent removed invacuo. The residue was purified by flash chromatography to yield4-[methanesulfonyl-(2-methoxy-ethyl)-amino]-piperidine-1-carboxylic acidtert-butyl ester (0.474 g). Treatment of this compound with HCl inDCM/MeOH yielded the desired amine, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.78 (2H, m), 1.92 (2H, m), 2.21 (2H, t, J=10.9Hz), 2.90 (3H, s), 3.07 (2H, br d, J=11.6 Hz), 3.38 (5H, m), 3.54 (2H,t, J=6.3 Hz), 3.68 (1H, m), 8.83 (2H, s), 3.94 (4H, m), 4.10 (4H, m),7.38 (1H, s), 7.50 (1H, t, J=7.7 Hz), 7.60 (1H, d, J=8.2 Hz), 8.29 (1H,d, J=7.1 Hz), 9.02 (1H, s), 10.10 (1H, br s); MS (ESI⁺) 586 (MH⁺).

1382-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-propan-2-ol

Via2-[1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-propan-2-ol,prepared from 2-piperidin-4-yl-propan-2-ol.

Amine preparation: To a solution of piperidine-4-carboxylic acid ethylester (3.0 g) stirring in dry MeCN (30 mL) was added potassium carbonate(2.90 g) followed by benzyl bromide (2.5 mL). The mixture was warmed to78° C. After 3 h the mixture was cooled, poured into water (100 mL) andextracted into ethyl acetate, dried (MgSO₄) and the solvent removed invacuo. The residue was purified using flash chromatography to give1-benzyl-piperidine-4-carboxylic acid ethyl ester (2.17 g) as a paleyellow oil.

To a solution of 1-benzyl-piperidine-4-carboxylic acid ethyl ester (1.0g) stirring in dry THF (15 mL) under a nitrogen atmosphere at 0° C., wasadded methyl magnesium bromide (3.0M solution in diethyl ether; 8.10 mL)and stirred for 2 h at 0° C. The reaction mixture was quenched withsaturated ammonium chloride solution and extracted into ethyl acetate,dried (MgSO₄) and the solvents removed in vacuo to give2-(1-benzyl-piperidine-4-yl)-propan-2-ol (1.11 g) as a white solid.

To a suspension of 10% palladium on charcoal (40 mg) stirring in dryMeOH (10 mL) under a nitrogen atmosphere was added2-(1-benzyl-piperidine-4-yl)-propan-2-ol (0.25 g) and the mixture warmedto 60° C. After 3 h the mixture was cooled, filtered through a celitebed and the solvents removed in vacuo to yield the desired amine (153mg) as a colourless oil.

¹H NMR (400 MHz, d₆-DMSO) 1.03 (s, 6H), 1.09 (m, 1H), 1.23 (m, 2H), 1.66(m, 2H), 1.98 (m, 2H), 2.97 (m, 2H), 3.83 (m, CH₂×2+CH₂), 4.00 (m, 4H),7.46 (t, 2H, J=7.3 Hz), 7.65 (d, 1H, J=8.2 Hz), 8.21 (d, 1 h, J=7.1 Hz),8.87 (s, 1H), 13.15 (br s, 1H); MS (ESI⁺) 493.2 (MH⁺).

1392-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(1-oxy-pyridin-3-ylmethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine

Via2-chloro-4-morpholin-4-yl-6-[4-(1-oxy-pyridin-3-ylmethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine,prepared from 1-(1-oxy-pyridin-3-ylmethyl)-piperazine.

Amine preparation: A mixture of 3-pyridylcarbinol-N-oxide (2.09 g) andthionyl chloride (1.65 mL) in dry chloroform (20 mL) was heated at 60°C. After 3 h, excess thionyl chloride was decomposed by addition ofethanol (0.5 mL). The resulting mixture was concentrated in vacuo.Trituration with ether/acetone gave 3-chloromethyl-pyridine 1-oxide(2.27 g). A mixture of 3-chloromethyl-pyridine 1-oxide (1.25 g),1-BOC-piperazine (1.47 g), K₂CO₃ (1.23 g) was heated at reflux in dryMeCN (25 mL). After 18 h, the reaction mixture was cooled, diluted withwater, extracted with DCM, dried (Na₂SO₄), filtered and concentrated invacuo then purified by flash chromatography (4% MeOH/DCM) to give4-(1-oxy-pyridin-3-ylmethyl)-piperazine-1-carboxylic acid tert-butylester (1.18 g). Treatment of this compound with HCl in DCM/MeOH yieldedthe desired amine, which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 2.54 (m, 4H, 2×CH₂), 2.61 (m, 4H, 2×CH₂), 3.49(s, 2H, CH₂), 3.85 (s, 2H, CH₂), 3.90-3.93 (m, 4H, 2×CH₂), 4.07-4.10 (m,4H, 2×CH₂), 7.18-7.22 (m, 2H, 2×ArH), 7.37 (s, H, ArH), 7.49 (t, H, ArH,J=7.75 Hz), 8.11 (d, H, ArH, J=5.2 Hz), 8.27 (m, 2H, 2×ArH), 9.30 (s, H,ArH), 10.27 (br s, H, NH); MS (ESI⁺) 543.3 (MH⁺).

1402-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-morpholin-4-ylmethyl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine

Via2-chloro-4-morpholin-4-yl-6-(4-morpholin-4-ylmethyl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared from 4-piperidin-4-ylmethyl-morpholine.

Amine preparation: To a solution of 1-BOC-piperidine-4-ethyl ester (2.0g), stirring in dry DCM (30 mL), at −78° C. under a nitrogen atmospherewas added diisobutylaluminium hydride solution (1.0M in hexane; 8.0 mL),the mixture was stirred at −78° C. for 2 h, then warmed to roomtemperature and quenched with MeOH (1 mL). The mixture was extractedinto DCM and dried (MgSO₄), and the solvents removed in vacuo to give aresidue which was purified using flash chromatography to give1-BOC-4-formyl-piperidine (457 mg).

To a solution of 1-BOC-4-formyl-piperidine (210 mg) in dry1,2-dichloroethane (10 mL), was added morpholine (86 mg) and glacialacetic acid (60 μL) and stirred for 1 h at room temperature. To themixture was added sodium triacetoxyborohydride (272 mg) and the mixturestirred for 12 h. The reaction mixture was extracted into DCM (25 mL),washed with 50% sodium bicarbonate solution (10 mL), brine (10 mL), anddried (MgSO₄). The solvents were removed in vacuo to give a residuewhich was purified by flash chromatography to give4-morpholine-4-ylmethyl-piperidine-1-carboxylic acid tert-butyl ester(120 mg). Treatment with TFA yielded the desired amine which wasisolated as the TFA salt.

¹H NMR (400 MHz, d₆-DMSO) 1.13 (m, 2H), 1.50 (m, 1H), 1.67 (m, 2H), 2.04(m, 2H), 2.10 (d, 2H, J=7.2 Hz), 2.28 (br s, 4H), 2.89 (m, 2H), 3.53 (m,4H), 3.77 (m, 4H+CH₂), 3.99 (m, 4H), 7.45 (t, 2H, J=9.8 Hz), 7.65 (d,1H, J=8.2 Hz), 8.21 (d, 1H, J=6.8 Hz), 8.87 (s, 1H), 13.15 (bs, 1H); MS(ESI⁺) 534.3 (MH⁺).

141{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-ylmethyl}-(2-methoxy-ethyl)-methyl-amine

Via[1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-ylmethyl]-(2-methoxy-ethyl)-methyl-amine,prepared from (2-methoxy-ethyl)-methyl-piperidin-4-ylmethyl-amine. Theamine, (2-methoxy-ethyl)-methyl-piperidin-4-ylmethylamine, was preparedin an analogous manner to that for 140 usingN-(2-methoxyethyl)methylamine.

¹H NMR (400 MHz, d₆-DMSO) 1.09 (m, CH₂), 1.44 (m, CH₂) 1.67 (m, CH₂),2.04 (t, CH₂, J=11.5 Hz), 2.15 (m, 3H+CH₂), 2.43 (t, CH₂, J=6.0 Hz),2.90 (m, CH₂), 3.21 (s, 3H), 3.39 (m, CH₂), 3.83 (m, 4H+CH₂), 3.99 (m,4H), 7.46 (t, 1H, J=6.2 Hz), 7.66 (d, 1H, J=8.3 Hz), 8.21 (d, 1H, J=7.4Hz), 8.87 (s, 1H), 13.25 (br s, 1H); MS (ESI⁺) 536.4 (MH⁺).

142{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-ylmethyl}-dimethyl-amine

Via[1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-ylmethyl]-dimethyl-amine,prepared from dimethyl-piperidin-4-ylmethyl-amine. The amine,dimethyl-piperidin-4-ylmethyl-amine, was prepared in an analogous mannerto that for 140 using dimethylamine.

¹H NMR (400 MHz, d₆-DMSO): 1.11 (m, CH₂), 1.42 (m, CH), 1.66 (m, CH₂),2.03 (m, CH₂×2), 2.08 (s, 6H), 2.89 (m, 2H), 3.82 (m, 4 h, +CH₂), 3.99(m, 4H), 7.45 (t, 2H, J=7.3 Hz), 7.65 (d, 1H, J=8.3 Hz), 8.21 (d, 1H,J=7.3 Hz), 8.87 (s, 1H), 13.2 (br s, 1H); MS (ESI⁺) 492.3 (MH⁺).

143{1-[2-(H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-3-yl}-(2-methoxy-ethyl)-methyl-amine

Via[1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-3-yl]-(2-methoxy-ethyl)-methyl-amine,prepared from (2-methoxy-ethyl)-methyl-piperidin-3-yl-amine.

Amine preparation: A mixture of 1-BOC-3-piperidone (0.50 g),N-(2-methoxyethyl)methylamine (0.29 g), sodium triacetoxyborohydride(0.74 g) and acetic acid (0.14 mL) in 1,2-dichloroethane (30 mL) wasstirred at room temperature for 6 h. The reaction mixture was thendiluted with DCM, washed with sodium bicarbonate solution, dried (MgSO₄)and the solvent removed in vacuo. The residue was purified by flashchromatography to yield3-[(2-methoxy-ethyl)-methyl-amino]-piperidine-1-carboxylic acidtert-butyl ester (0.556 g). Treatment of this compound with HCl inDCM/MeOH yielded the desired amine, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 1.27 (1H, m), 1.60 (1H, m), 1.77 (1H, m), 1.98(3H, m), 2.35 (3H, s), 2.68 (3H, m), 2.92 (1H, d, J=10.5 Hz), 3.10 (1H,m), 3.34 (3H, s), 3.44 (2H, m), 3.86 (2H, s), 3.92 (4H, m), 4.08 (4H,m), 7.37 (1H, s), 7.50 (1H, t, J=7.7 Hz), 7.60 (1H, d, J=8.3 Hz), 8.27(1H, d, J=7.3 Hz), 9.01 (1H, s), 10.20 (1H, br s); MS (ESI⁺) 522 (MH⁺).

1441-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-3-carboxylicacid methylamide

Via1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidine-3-carboxylicacid methylamide, prepared from piperidine-3-carboxylic acidmethylamide.

Amine preparation: To a solution of1-(tert-butoxycarbonyl)-3-piperidine-carboxylic acid (0.50 g) in DMF (3mL) was added 1,1′-carbonyldiimidazole (0.70 g). The reaction mixturewas stirred for 4 h and then triethylamine (0.60 mL) followed bymethylamine hydrochloride (0.29 g) was added. After stirring for 24 hthe reaction mixture was then diluted with ethyl acetate, washed withwater, brine, dried (MgSO₄) and the solvent removed in vacuo. Theresidue was purified by flash chromatography to yield3-methylcarbamoyl-piperidine-1-carboxylic acid tert-butyl ester (0.41g). Treatment of this compound with HCl in DCM/MeOH yielded the desiredamine, which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.58-1.94 (4H, m), 2.50 (2H, m), 2.65 (2H, m),2.84 (3H, d), 2.80 (1H, m), 3.85 (2H, m), 3.92 (4H, m), 4.10 (4H, m),7.00 (1H, br s), 7.40 (1H, s), 7.50 (1H, t, J=7.7 Hz), 7.60 (1H, d,J=8.2 Hz), 8.29 (1H, d, J=7.1 Hz), 9.03 (1H, s), 10.10 (1H, br s); MS(ESI⁺) 492 (MH⁺).

1452-(1H-Indazol-4-yl)-6-(3-methoxymethyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-Chloro-6-(3-methoxymethyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 3-methoxymethyl-piperidine.

Amine preparation: To a solution of 1-BOC-3-piperidine carboxylic acid(3.0 g) in DMF (25 mL) was added potassium carbonate (3.62 g) followedby iodomethane (4.07 mL). After 2.5 h the reaction was diluted withwater and extracted into diethyl ether. The organic layer was washedwith brine, separated and dried (MgSO₄). The solvent was evaporated togive piperidine-1,3-dicarboxylic acid 1-tert-butyl ester 3-methyl ester(2.98 g).

To a suspension of lithium aluminium hydride (1.41 g) in THF (15 mL) at0° C. was added piperidine-1,3-dicarboxylic acid 1-tert-butyl ester3-methyl ester as a solution in THF (10 mL) and the mixture stirred atroom temperature for 3 h. The reaction was cooled to 0° C. and quenchedby addition of aqueous ammonium chloride and the mixture filteredthrough celite. The filtrate was diluted with ethyl acetate, washed withwater, separated and dried (MgSO₄). The solvent was evaporated to give3-hydroxymethyl-piperidine-1-carboxylic acid tert-butyl ester (1.25 g).To a solution of this alcohol (422 mg) in THF (8 mL) was added sodiumhydride (94 mg; 60% dispersion in mineral oil). After 15 min iodomethane(0.49 mL) was added and the reaction stirred for 18 h. The mixture wasthen diluted with ethyl acetate and washed sequentially with water,brine, separated and dried (MgSO₄). The solvent was evaporated and theresidue purified by flash chromatography to give3-methoxymethyl-piperidine-1-carboxylic acid tert-butyl ester (414 mg).To a solution of 3-methoxymethyl-piperidine-1-carboxylic acid tert-butylester (204 mg) in DCM (3 mL) was added 2M hydrogen chloride in diethylether and the mixture stirred for 18 h. The solvent was evaporated togive the desired amine (168 mg).

¹H NMR (400 MHz, CDCl₃) 1.06 (s, 1H), 1.63-1.75 (m, 2H), 1.90-1.95 (m,3H), 2.12 (m, 1H), 2.88 (m, 1H), 2.99 (m, 1H), 3.25 (m, 2H), 3.31 (s,3H), 3.83 (s, 2H), 3.92 (t, J=4.8 Hz, 4H), 4.09 (t, J=4.8 Hz, 4H), 7.35(s, 1H), 7.50 (t, J=7.8 Hz, 1H), 7.57 (d, J=8.2 Hz, 1H), 8.28 (d, J=7.4Hz, 1H), 9.02 (s, 1H), 10.10 (br s, 1H); MS (ESI⁺) 479.26 (MH⁺).

1462-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-2-ylmethyl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine

Via2-chloro-4-morpholin-4-yl-6-(4-pyridin-2-ylmethyl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared from 2-piperidin-4-ylmethyl-pyridine.

Amine preparation: 4-Pyridin-2-ylmethyl-piperidine-1-carboxylic acidtert-butyl ester was synthesized according to the procedure described inJ. Org. Chem. 2001, 66, 2487. It was treated further with HCl inDCM/MeOH to yield the desired amine.

¹H NMR (400 MHz, CDCl₃) 1.40-1.50 (2H, m), 1.64-1.70 (2H, m), 1.81-1.92(1H, m), 2.06-2.13 (2H, m), 2.74 (2H, d, J=7.2 Hz), 2.96-3.03 (2H, m),3.83 (2H, s), 3.90-3.96 (4H, m), 4.05 (4H, m), 7.10-7.13 (2H, m), 7.32(1H, s), 7.50-7.53 (1H, m), 7.56-7.62 (2H, m), 8.26 (1H, d, J=6.8 Hz),8.52-8.54 (1H, m), 9.01 (1H, s), 10.20 (1H, br); MS (ESI⁺) 526 (MH⁺).

1472-(1H-Indazol-4-yl)-6-[4-(2-methoxy-ethoxy)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-chloro-6-[4-(2-methoxy-ethoxy)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 4-(2-methoxy-ethoxy)-piperidine.

Amine preparation: To 4-hydroxy-piperidine-1-carboxylic acid tert-butylester (1.5 g) in dry DMF (15 mL) was added sodium hydride (447 mg).After stirring for 2 hrs at room temperature, 2-bromoethyl methyl ether(0.7 mL) was added. The reaction mixture was heated to 40° C. overnight,and then diluted with ethyl acetate, washed with water and dried(MgSO₄). The solvent was evaporated and the residue purified by flashchromatography to give 4-(2-methoxy-ethoxy)-piperidine-1-carboxylic acidtert-butyl ester (447 mg). Treatment of this compound with HCl inDCM/MeOH yielded the desired amine, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.72 (m, 2H), 1.94 (m, 2H), 2.30 (m, 2H), 2.85(m, 2H), 3.40 (s, 4H), 3.55 (m, 2H), 3.61 (m, 2H), 3.84 (s, 2H), 3.92(m, 4H), 4.09 (m, 4H), 7.35 (s, 1H), 7.52 (m, 1H), 7.59 (d, J=8.2 Hz,1H), 8.28 (d, J=6.8 Hz, 1H), 9.02 (s, 1H); MS (ESI⁺) 509 (MH⁺).

1486-((3R,5S)-3,5-Dimethyl-4-thiazol-2-ylmethyl-piperazin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-chloro-6-((3R,5S)-3,5-dimethyl-4-thiazol-2-ylmethyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from (2R,6S)-2,6-dimethyl-1-thiazol-2-ylmethyl-piperazine.

Amine preparation: 2-Thiazolecarboxaldehyde was converted to thecorresponding alcohol by treatment with sodium borohydride. Reactionwith methanesulfonyl chloride yielded methanesulfonic acidthiazol-2-ylmethyl ester. A mixture of methanesulfonic acidthiazol-2-ylmethyl ester (393 mg),(3R,5S)-3,5-dimethyl-piperazine-1-carboxylic acid tert-butyl ester(described previously, 435 mg), potassium carbonate (309 mg) andtetrabutyl ammonium iodide (827 mg) was heated to reflux in MeCN (10mL). After 4 days the reaction mixture was cooled, diluted with ethylacetate, washed with water and dried (MgSO₄). The solvent was evaporatedand the residue purified by flash chromatography to give(R)-3,5-dimethyl-4-thiazol-2-ylmethyl-piperazine-1-carboxylic acid(S)-tert-butyl ester (314 mg). Treatment of this compound with HCl inDCM/MeOH yielded the desired amine, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.12 (d, J=6.0 Hz, 6H), 2.06 (m, 2H), 2.85 (m,4H), 3.80 (s, 2H), 3.92 (m, 4H), 4.09 (m, 4H), 4.17 (s, 2H), 7.24 (d,J=3.2 Hz, 1H), 7.38 (s, 1H), 7.51 (m, 1H), 7.59 (d, J=8.3 Hz, 1H), 7.74(d, J=3.3 Hz, 1H), 8.29 (d, J=7.3 Hz, 1H), 9.03 (s, 1H), 10.1 (br s,1H); MS (ESI⁺) 561 (MH⁺).

1492-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(1-oxy-pyridin-2-ylmethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine

Via2-chloro-4-morpholin-4-yl-6-[4-(1-oxy-pyridin-2-ylmethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine,prepared from 1-(1-oxy-pyridin-2-ylmethyl)-piperazine.

Amine preparation: To 2-pyridylcarbinol (2.26 g) in chloroform (20 mL)was added mCPBA (5.57 g) at 0° C. The reaction mixture was allowed tostir at that temperature for 1 h, the solvent was removed in vacuo andthe residue was purified using flash chromatography to yield(1-oxy-pyridin-2-yl)-methanol as a white solid (2.73 g). To a precooled(0° C.) solution of (1-oxy-pyridin-2-yl)-methanol (2.73 g) in drychloroform (25 mL) was added thionyl chloride (2.07 mL) dropwise. Thereaction mixture was heated at 60° C. for 3 h, cooled to roomtemperature, quenched with ethanol (1 mL and reduced in vacuo. Theresidue was taken up in DCM, washed with aqueous sodium bicarbonate,dried (MgSO₄) and the solvent removed in vacuo to give2-chloromethyl-pyridine 1-oxide as an oil (2.60 g), which solidified onstanding. BOC-piperazine (500 mg) and 2-chloromethyl-pyridine 1-oxide(385 mg) were refluxed in MeCN (10 mL) with potassium carbonate (550mg). After 4 h, the reaction mixture was reduced in vacuo, the residuewas taken up in DCM, washed with brine, dried (MgSO₄) and the solventremoved in vacuo to give4-(1-oxy-pyridin-2-ylmethyl)-piperazine-1-carboxylic acid tert-butylester (788 mg). Treatment of this compound with HCl in DCM/MeOH yieldedthe desired amine, which was isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 2.65-2.76 (8H, m), 3.88 (2H, s), 3.89 (2H, s),3.90-3.92 (4H, m), 4.03-4.05 (4H, m), 7.14-7.18 (1H, m), 7.26-7.29 (1H,m), 7.37 (1H, s), 7.48-7.51 (1H, m), 7.55-7.59 (2H, m), 8.23-8.29 (2H,m), 9.02 (1H, s); MS (ESI⁺) 543 (MH⁺).

1502-(1H-Indazol-4-yl)-6-[4-(2-methoxy-ethyl)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-Chloro-6-[4-(2-methoxy-ethyl)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 4-(2-methoxy-ethyl)-piperidine.

Amine preparation: To a solution of 4-piperidine ethanol (540 mg) inMeCN (8 mL) was added triethylamine (0.70 mL) followed by tert-butyldicarbonate (1.00 g). The mixture was stirred at room temperature for 22h and partitioned between 0.5 M hydrochloric acid and ethyl acetate. Theorganic layer was washed with brine, dried (MgSO₄) and the solventevaporated to give 4-(2-hydroxy-ethyl)-piperidine-1-carboxylic acidtert-butyl ester (897 mg). This was dissolved in THF (14 mL) and sodiumhydride added (172 mg; 60% dispersion in mineral oil) at 0° C. After 30min iodomethane (0.97 mL) was added and the mixture stirred for 22 h.The mixture was diluted with ethyl acetate and washed with water, brineand dried (MgSO₄). The solvent was evaporated and the residue purifiedby flash chromatography to give4-(2-methoxy-ethyl)-piperidine-1-carboxylic acid tert-butyl ester (724mg). To a solution of 4-(2-methoxy-ethyl)-piperidine-1-carboxylic acidtert-butyl ester (284 mg) in DCM (5 mL) was added 2M hydrogen chloridein diethyl ether (5.0 mL) and the mixture stirred for 21 h. The solventwas evaporated to give the desired amine as a white solid.

¹H NMR (400 MHz, CDCl₃) 1.45-1.25 (m, 5H), 1.71 (m, 2H), 2.10 (m, 2H),2.98 (m, 2H), 3.33 (s, 3H), 3.42 (t, J=6.5 Hz, 2H), 3.83 (s, 2H), 3.92(t, J=4.8 Hz, 4H), 4.09 (t, J=4.8 Hz, 4H), 7.36 (s, 1H), 7.51 (t, J=7.7Hz, 1H), 7.59 (d, J=8.3 Hz, 1H), 8.28 (d, J=6.7 Hz, 1H), 9.02 (s, 1H),10.05 (br s, 1H); MS (ESI⁺) 493.27 (MH⁺).

1512-(1H-Indazol-4-yl)-6-(4-methanesulfonyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-chloro-6-(4-methanesulfonyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 4-methanesulfonyl-piperidine.

Amine preparation: A mixture of4-methanesulfonyloxy-piperidine-1-carboxylic acid tert-butyl ester(1.015 g), sodium thiomethoxide (635 mg) was heated to 80° C. in DMF (10mL). After 4 h, the reaction mixture was diluted with water, extractedwith ethyl acetate, dried (MgSO₄), filtered and concentrated in vacuoand then purified by flash chromatography to give4-methylsulfanyl-piperidine-1-carboxylic acid tert-butyl ester (600 mg).To a solution of 4-methylsulfanyl-piperidine-1-carboxylic acidtert-butyl ester (600 mg) in chloroform (15 mL) was added mCPBA (1.46g). After stirring for 2 days, the reaction mixture was diluted withDCM, washed with sodium bicarbonate solution, dried (MgSO₄) and thesolvent removed in vacuo to yield4-methanesulfonyl-piperidine-1-carboxylic acid tert-butyl ester (505 mg)as a white solid. Treatment of this compound with HCl in DCM/MeOHyielded the desired amine, isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.90-2.05 (2H, m), 2.10-2.18 (4H, m), 2.90 (3H,s), 2.91-2.94 (1H, m), 3.21 (2H, d), 3.88 (2H, s), 3.91-3.93 (4H, m),4.10-4.12 (4H, m), 7.40 (1H, s), 7.48-7.52 (1H, m), 7.58 (1H, d), 8.25(1H, d), 9.05 (1H, s), 10.25 (s br, 1H); MS (ESI⁺) 513 (MH⁺).

152{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(3-methanesulfonyl-propyl)-methyl-amine

Via[1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-(3-methanesulfonyl-propyl)-methyl-amine,prepared from (3-methanesulfonyl-propyl)-methyl-piperidin-4-yl-amine.

Amine preparation: Toluene-4-sulfonic acid 3-methylsulfanyl-propyl esterwas prepared from 3-(methylthio)-1-propanol using standard conditions.Treatment with mCPBA in DCM yielded toluene-4-sulfonic acid3-methanesulfonyl-propyl ester. A mixture of4-methylamino-piperidine-1-carboxylic acid tert-butyl ester andtoluene-4-sulfonic acid 3-methanesulfonyl-propyl ester was heated inMeCN in the prescence of potassium carbonate to yield4-[(3-methanesulfonyl-propyl)-methyl-amino]-piperidine-1-carboxylic acidtert-butyl ester. Treatment of this compound with HCl in DCM/MeOHyielded the desired amine, isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.50-1.70 (m, 4H, 2×CH₂), 1.90-1.97 (m, 2H,CH₂), 2.00-2.05 (m, 2H, CH₂), 2.21 (s, 3H, CH₃), 2.38 (m, H, CH), 2.55(m, 2H, CH₂), 2.74 (s, 3H, CH₂), 2.96-3.04 (m, 4H, 2×CH₂), 3.75 (s, 2H,CH₂), 3.83-3.89 (m, 4H, 2×CH₂), 4.00-4.02 (m, 4H, 2×CH₂), 7.28 (s, H,CH), 7.41 (t, H, ArH, J=7.74 Hz), 7.50 (d, H, ArH, J=8.24 Hz), 8.18 (d,H, ArH, J=7.05 Hz), 8.93 (s, H, ArH); MS (ESI⁺) 584.39 (MH⁺).

1532-(1H-Indazol-4-yl)-6-[4-(3-methoxy-propane-1-sulfonyl)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-chloro-6-[4-(3-methoxy-propane-1-sulfonyl)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 4-(3-methoxy-propane-1-sulfonyl)-piperidine.

Amine preparation: A mixture of4-methanesulfonyloxy-piperidine-1-carboxylic acid tert-butyl ester (2.82g) and potassium thioacetate (2.31 g) was heated to 60° C. in DMF (10mL). After 4 h, the reaction mixture was diluted with water, extractedwith ethyl acetate, dried (MgSO₄), filtered and concentrated in vacuoand then purified by flash chromatography to give4-acetylsulfanyl-piperidine-1-carboxylic acid tert-butyl ester (1.80 g).To a solution of 4-acetylsulfanyl-piperidine-1-carboxylic acidtert-butyl ester (607 mg) in dry MeOH (5 mL) was added a solution ofsodium methoxide in MeOH (25% wt. 0.59 mL). After stirring for 15 min,toluene-4-sulfonic acid 3-methoxy-propyl ester (described elsewhere, 571mg) was added in MeOH. After 24 h, the reaction mixture was diluted withwater, extracted with ethyl acetate, dried (MgSO₄), filtered andconcentrated in vacuo to yield4-(3-methoxy-propylsulfanyl)-piperidine-1-carboxylic acid tert-butylester. Treatment with mCPBA (as described above) yielded4-(3-methoxy-propane-1-sulfonyl)-piperidine-1-carboxylic acid tert-butylester. Treatment of this compound with HCl in DCM/MeOH yielded thedesired amine, isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 1.95-2.20 (8H, m), 2.85-2.91 (1H, m), 3.02-3.06(2H, t), 3.13-3.20 (2H, d), 3.31 (3H, s), 3.52 (2H, t), 3.88 (2H, s),3.94-3.99 (4H, m), 4.09-4.13 (4H, m), 7.35 (1H, s), 7.45-7.52 (1H, m),7.55 (1H, d), 8.29 (1H, d), 9.05 (1H, s), 10.30 (1H, br s); MS (ESI⁺)571 (MH⁺).

154(R)-1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-3-carboxylicacid methylamide

Via(R)-1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidine-3-carboxylicacid methylamide, prepared from (R)-piperidine-3-carboxylic acidmethylamide.

Amine preparation: To a suspension of (S)-(−)-nipecotic acidhydrochloride (1.0 g) in MeCN (10 mL) was added triethylamine (1.85 mL)followed by di-tert-butyldicarbonate (1.45 g). After stirring for 24 hthe reaction mixture was then diluted with DCM, washed with brine, dried(MgSO₄) and the solvent removed in vacuo. The residue was purified byflash chromatography to yield piperidine-1,3-dicarboxylic acid1-tert-butyl ester (0.54 g). Procedure continued as for 144.

¹H NMR (400 MHz, CDCl₃) 1.58-1.94 (4H, m), 2.50 (2H, m), 2.65 (2H, m),2.84 (3H, d), 2.80 (1H, m), 3.85 (2H, m), 3.92 (4H, m), 4.10 (4H, m),7.00 (1H, br s), 7.40 (1H, s), 7.50 (1H, t, J=7.7 Hz), 7.60 (1H, d,J=8.2 Hz), 8.29 (1H, d, J=7.1 Hz), 9.03 (1H, s), 10.10 (1H, br s); MS(ESI⁺) 492 (MH⁺).

155(S)-1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-3-carboxylicacid methylamide

Via(S)-1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidine-3-carboxylicacid methylamide, prepared from (S)-piperidine-3-carboxylic acidmethylamide. Amine preparation as for 154, utilizing (R)-(+)-nipecoticacid hydrochloride as the starting material.

¹H NMR (400 MHz, CDCl₃): 1.58-1.94 (4H, m), 2.50 (2H, m), 2.65 (2H, m),2.84 (3H, d), 2.80 (1H, m), 3.85 (2H, m), 3.92 (4H, m), 4.10 (4H, m),7.00 (1H, br s), 7.40 (1H, s), 7.50 (1H, t, J=7.7 Hz), 7.60 (1H, d,J=8.2 Hz), 8.29 (1H, d, J=7.1 Hz), 9.03 (1H, s), 10.10 (1H, br s); MS(ESI⁺) 492 (MH⁺).

1566-(4-Imidazol-1-ylmethyl-piperidin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3.2-d]pyrimidine

Via2-chloro-6-(4-imidazol-1-ylmethyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 4-imidazol-1-ylmethyl-piperidine.

Amine preparation: To a solution of4-hydroxymethyl-piperidine-1-carboxylic acid tert-butyl ester (250 mg),in dry THF (15 mL), was added carbon tetrabromide (769 mg), andtriphenyl phosphine (609 mg). The reaction mixture was stirred at roomtemperature for 24 h, and then the solvents were evaporated in vacuo togive a residue which was purified by flash chromatography to give4-bromomethyl-piperidine-1-carboxylic acid tert-butyl ester (279 mg), asa colourless oil. To a solution of 4-bromomethyl-piperidine-1-carboxylicacid tert-butyl ester (240 mg), in dry DMF (5.0 mL), was added imidazole(129 mg). The reaction mixture was heated in a sealed reaction vial at100° C. for 24 h, then cooled and the contents evaporated onto flashsilica for purification. Treatment of this compound with HCl in DCM/MeOHyielded the desired amine, isolated as the hydrochloride salt.

¹H NMR (400 MHz, d₆-DMSO) 1.23 (m, 2H), 1.47 (m, 2H), 1.69 (m, CH), 2.01(m, 2H), 2.92 (m, 2H), 3.83 (m, 4H+CH₂×2), 3.99 (m, 4H), 6.87 (s, 1H),7.13 (s, 1H), 7.46 (t, 1H, J=7.6 Hz), 7.58 (s, 1H), 7.65 (d, 1H, J=8.2Hz), 8.21 (d, 1H, J=7.3 Hz), 8.87 (s, 1H), 13.2 (br s, 1H); MS (ESI⁺)515.2 (MH⁺).

1572-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-morpholin-4-ylmethyl-thieno[3,2-d]pyrimidine

Via2-Chloro-4-morpholin-4-yl-6-morpholin-4-ylmethyl-thieno[3,2-d]pyrimidine,prepared from morpholine.

¹H NMR (400 MHz, CDCl₃) 2.58-2.63 (4H, m), 3.72-3.78 (4H, m), 3.82 (2H,s), 3.88-3.93 (4H, m), 4.05-4.11 (4H, m), 7.38 (1H, s), 7.50-7.55 (2H,m), 7.60 (1H, d, J=8.3 Hz), 8.28 (1H, d, J=7.2 Hz), 9.00 (1H, s), 10.10(1H, br); MS (ESI⁺) 437 (MH⁺).

1582-(1H-Indazol-4-yl)-6-(3-methyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine.

Via2-Chloro-6-(3-methyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 3-methyl-piperidine.

¹H NMR (400 MHz, CDCl₃) 0.88 (4H, m), 1.70 (5H, m), 2.04 (1H, m), 2.90(2H, m), 3.80 (2H, s), 3.90 (4H, m), 4.10 (4H, m), 7.36 (1H, s), 7.50(1H, t, J=7.7 Hz), 7.58 (1H, d, J=8.2 Hz), 8.29 (1H, d, J=7.1 Hz), 9.02(1H, s), 10.10 (1H, br s); MS (ESI⁺) 490 (MH⁺).

159{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-3-yl}-methanol

Via[1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-3-yl]-methanol,prepared from piperidin-3-yl-methanol.

¹H NMR (400 MHz, CDCl₃) 1.18 (m, 1H), 1.58-1.88 (m, 4H), 2.12 (m, 1H),2.27 (m, 1H), 2.77 (m, 1H), 2.93 (m, 1H), 3.56 (dd, J=10.6 Hz, 6.4 Hz,1H), 3.64 (dd, J=10.6 Hz, 5.5 Hz, 1H), 3.84 (s, 2H), 3.92 (t, J=4.8 Hz,4H), 4.09 (t, J=4.8 Hz, 4H), 7.36 (s, 1H), 7.51 (t, J=7.7 Hz, 1H), 7.59(d, J=8.3 Hz, 1H), 8.27 (d, J=7.3 Hz, 1H), 9.01 (s, 1H), 10.10 (br s,1H); MS (ESI⁺) 465.20 (MH⁺).

1602-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-ethanol

Via2-[1-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperidin-4-yl]-ethanol,prepared from 2-piperidin-4-yl-ethanol.

¹H NMR (400 MHz, d₆-DMSO) 1.18 (m, 2H), 1.36 (m, 3H), 1.63 (m, 2H), 2.02(m, 2H), 2.91 (m, 2H), 3.30-3.45 (m, 2H), 3.84 (m, 6H), 3.99 (t, J=4.8Hz, 4H), 4.35 (t, J=4.8 Hz, 4H), 7.46 (m, 2H), 7.65 (d, J=8.3 Hz, 1H),8.21 (d, J=7.8 Hz, 1H), 8.87 (s, 1H); MS (ESI⁺) 479.25 (MH⁺).

1611-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-4-thiazol-2-yl-piperidin-4-ol

Via1-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-4-thiazol-2-yl-piperidin-4-ol,prepared from 4-thiazol-2-yl-piperidin-4-ol.

¹H NMR (400 MHz, d₆-DMSO) 1.73 (m, 2H), 2.14 (m, 2H), 2.53 (m, 2H), 2.78(m, 2H), 3.83 (m, 4H), 3.91 (s, 2H), 4.01 (m, 4H), 5.95 (s, 1H), 7.46(t, 1H, J=7.6 Hz), 7.49 (s, 1H), 7.57 (d, 1H, J=3.1 Hz), 7.65 (d, 1H,J=8.3 Hz), 7.72 (d, 1H, J=3.3 Hz), 8.21 (d, 1H, J=7.3 Hz), 8.88 (s, 1H),13.18 (br s, 1H); MS (ESI⁺) 534.3 (MH⁺).

Preparation of Boronate Ester Monomers for the Synthesis of 162 and 163

A solution of4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indazole (400 mg) indry DMF (4 mL) precooled to 0° C. was added to a suspension of sodiumhydride (80 mg) in dry THF (5 mL) at −78° C. under inert atmosphere.After 30 min, iodomethane (112 uL) was added to the mixture at −78° C.The reaction mixture was allowed slowly to warm up to room temperatureovernight and then diluted with DCM, washed with brine, dried (MgSO₄)and the solvent removed in vacuo. The residue was purified using flashchromatography to yield1-methyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-1H-indazole(143 mg) and2-methyl-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-2H-indazole(160 mg).

1622-(1-Methyl-1H-indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Prepared in a similar manner to 59.

¹H NMR (400 MHz, CDCl₃) 2.32 (3H, s), 2.45-2.55 (4H, br m), 2.55-2.70(4H, br m), 3.83 (2H, s), 3.90-3.93 (4H, m), 4.03-4.06 (4H, m), 4.12(3H, s), 7.40 (1H, s), 7.49-7.52 (2H, m), 8.25-8.27 (1H, m), 8.90 (1H,s); MS (ESI⁺) 464 (MH⁺).

1632-(2-Methyl-2H-indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Prepared in a similar manner to 59.

¹H NMR (400 MHz, CDCl₃) 2.32 (3H, s), 2.45-2.55 (4H, br m), 2.55-2.70(4H, br m), 3.83 (2H, s), 3.90-3.93 (4H, m), 4.03-4.06 (4H, m), 4.30(3H, s), 7.48 (2H, m), 7.81 (1H, d, J=8.4 Hz), 8.28 (1H, d, J=6.8 Hz),8.90 (1H, s); MS (ESI⁺) 464 (MH⁺).

Alternative Synthetic Approaches 1642-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-thiazol-4-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine

Via2-chloro-4-morpholin-4-yl-6-(4-thiazol-4-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared from 1-thiazol-4-ylmethyl-piperazine, via2-chloro-4-morpholin-4-yl-6-(4-thiazol-4-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine,prepared according to the following procedure: to a suspension of4-thiazolecarboxylic acid (500 mg) in THF (10 mL) was addedborane-dimethylsulfide complex (0.73 mL). After 24 h the mixture wascooled to 0° C. and quenched by the addition of 2M hydrochloric acid andextracted into ethyl acetate. The organic layers were washed with brineand dried (MgSO₄). The solvent was evaporated and the residue stirred inDCM/MeOH overnight. The mixture was concentrated and the residuepurified by flash chromatography to give thiazol-4-yl-methanol (173 mg).To a solution of thiazol-4-yl-methanol (168 mg) in DCM (5 mL) was addedtriethylamine (0.33 mL) followed by methanesulfonyl chloride (0.17 mL)at 0° C. The mixture was stirred at room temperature for 10 min anddiluted with DCM, washed with brine and dried (MgSO₄). The crude productwas purified by flash chromatography to give methanesulfonic acidthiazol-4-yl methyl ester (263 mg). To a solution of2-chloro-4-morpholin-4-yl-6-piperazin-1-ylmethyl-thieno[3,2-d]pyrimidine(300 mg) and methanesulfonic acid thiazol-4-yl methyl ester (213 mg) inMeCN (10 mL) was added potassium carbonate (164 mg) and the mixtureheated at 80° C. for 8 h. The cooled mixture was filtered, the solventevaporated and the residue partitioned between DCM and water. Theorganic layer was washed with brine, dried (MgSO₄) and the solventevaporated. The residue was purified by flash chromatography to give thedesired product (249 mg).

¹H NMR (400 MHz, CDCl₃) 2.57 (br s, 4H), 3.71 (s, 2H), 3.79 (s, 2H),3.85 (t, J=4.8 Hz, 4H), 4.02 (t, J=4.8 Hz, 4H), 7.13 (s, 1H), 7.30 (s,1H), 7.43 (t, J=7.8 Hz, 1H), 7.51 (d, J=8.3 Hz, 1H), 8.21 (d, J=6.9 Hz,1H), 8.71 (d, J=2.0 Hz, 1H), 8.95 (s, 1H), 10.10 (br s, 1H); MS (ESI⁺)533.25 (MH⁺).

1651-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-3-phenoxy-propan-2-ol

Via4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-piperazine-1-carboxylicacid tert-butyl ester, prepared from 1-BOC-piperazine.

Treatment of4-[2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester with HCl in DCM/MeOH yielded2-(1H-indazol-4-yl)-4-morpholin-4-yl-6-piperazin-1-ylmethyl-thieno[3,2-d]pyrimidine.A mixture of2-(1H-indazol-4-yl)-4-morpholin-4-yl-6-piperazin-1-ylmethyl-thieno[3,2-d]pyrimidine(150 mg) and 1,2-epoxy-3-phenoxypropane (40 mg) was suspended in 50%DMF/0.1M sodium phosphate buffer (2 mL) and shaken at 55° C. overnight.After stirring for 24 h the reaction mixture was then diluted with DCM,washed with brine, dried (MgSO₄) and the solvent removed in vacuo. Theresidue was purified by flash chromatography.

¹H NMR (400 MHz, CDCl₃) 2.50-2.70 (8H, br), 2.70-2.82 (2H, br), 3.85(2H, s), 3.90-3.93 (4H, m), 3.99-4.01 (2H, m), 4.07-4.14 (5H, m),6.90-6.98 (3H, m), 7.26-7.30 (2H, m), 7.40 (1H, s), 7.50-7.53 (1H, m),7.59 (1H, d, J=8.3 Hz), 8.29 (1H, d, J=7.0 Hz), 9.02 (1H, s); MS (ESI⁺)568 (MH⁺).

1666-[4-(1H-Imidazol-2-ylmethyl)-piperazin-1-ylmethyl]-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

ia2-chloro-4-morpholin-4-yl-6-piperazin-1-ylmethyl-thieno[3,2-d]pyrimidine,prepared from 1-BOC-piperazine followed by treatment with HCl andsubsequent reaction with imidazole-2-carboxaldehyde using standardreductive amination conditions. The amine,2-chloro-6-[4-(1H-imidazol-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidinewas subsequently isolated.

¹H NMR (400 MHz, CDCl₃): 2.55-2.72 (8H, br m), 3.71 (2H, s), 3.89 (2H,s), 3.92-3.96 (4H, m), 4.03-4.11 (4H, m), 7.02 (2H, s), 7.32 (1H, s),7.51 (1H, t, J=8.0 Hz), 7.59 (1H, d, J=8.3), 8.29 (1H, d, J=6.6 Hz),9.03 (1H, s), 10.10 (1H, br m); MS (ESI⁺) 516 (MH⁺).

1676-[4-(3H-Imidazol-4-ylmethyl)-piperazin-1-ylmethyl]-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

In a similar manner to 167 (PI1343)2-chloro-6-[4-(3H-imidazol-4-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidinewas prepared by reductive amination with 5-formyl-imidazole-1-carboxylicacid tert-butyl ester followed by removal of the BOC group using HCl.

¹H NMR (400 MHz, d₆-DMSO) 2.45 (br s, 4H), 3.29 (s, 2H), 3.42 (br s,4H), 3.84 (m, 6H), 3.99 (t, 4H), 7.46 (m, 3H), 7.47 (s, 1H), 7.65 (d,J=7.4 Hz, 1H), 8.21 (d, J=7.4 Hz, 1H), 8.86 (s, 1H), 13.20 (s, 1H). MS(ESI⁺) 516 (MH⁺).

Synthesis Via Alkyl Bromide (Route 3)

1732-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-((2S,6R)-2,4,6-trimethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine

To (66) (1.5 g) in ethanol (30 mL) was added sodium borohydride (1 g).After 4 h the reaction mixture was quenched with brine and the resultingsolid was collected by filtration, and air dried to furnish 168(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-yl)-methanol (1.42g).

To a solution of 168 (1.42 g) in toluene (14 ml), warmed to 40° C., wasadded phosphorous tribromide (0.16 ml). The resulting mixture was thenheated to 100° C. for 6 h, was cooled, diluted with chloroform, washedwith brine, and dried (MgSO₄). The solvent was removed in vacuo to yield6-bromomethyl-2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidine, 169(1.40 g).

A mixture of (3R,5S)-3,5-dimethyl-piperazine-1-carboxylic acidtert-butyl ester (0.92 g),6-bromomethyl-2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidine (1 g)and potassium carbonate (1.59 g) in MeCN (10 ml) was heated to refluxfor 5 days. The reaction mixture was subsequently cooled, diluted withchloroform, washed with brine and dried (MgSO₄), and the solvent wasremoved in vacuo. The residue was purified by flash columnchromatography to yield(3S,5R)-4-(2-chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-3,5-dimethyl-piperazine-1-carboxylicacid tert-butyl ester, 170 (1.2 g). Treatment of this compound with HClin DCM/MeOH produced2-chloro-6-(2S,6R)-2,6-dimethyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,171, which was then methylated using 37% formaldehyde solution andsodium borohydride in MeOH, furnishing 172.

¹H NMR (400 MHz, CDCl₃) 1.17 (6H, d), 1.92 (2H, t), 2.3 (3H, s), 2.73(2H, d), 2.83 (2H, m), 3.95 (4H, m), 4.03 (4H, m), 4.18 (2H, s), 7.36(1H, s), 7.48 (1H, t), 7.56 (1H, d), 8.26 (1H, d), 9.00 (1H, s), 10.40(1H, br m); MS (ESI⁺) 478 (MH⁺).

174{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-1-methanesulfonyl-piperazin-2-yl}-methanol

Via[4-(2-Chloro-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl)-1-methanesulfonyl-piperazin-2-yl]-methanol,prepared from (1-methanesulfonyl-piperazin-2-yl)-methanol.

Amine preparation: To a suspension of piperazine-2-carboxylic aciddihydrochloride (10.0 g) in 1,4-dioxane (100 mL) and water (50 mL),cooled down to 0° C., was slowly added 17M sodium hydroxide solution(8.6 mL) followed by di-tert-butyldicarbonate (11.8 g). The reactionmixture was warmed to room temperature and stirred for 5 h.Triethylamine (13.7 mL) followed by methanesulfonyl chloride (3.8 mL)was then added. After stirring for 24 h the reaction mixture wasconcentrated down, the residue diluted with ethyl acetate, washed with2M hydrochloric acid, brine, dried (MgSO₄) and the solvent removed invacuo to yield crude 4-methanesulfonyl-piperazine-1,3-dicarboxylic acid1-tert-butyl ester (8.46 g). This was subsequently dissolved in DMF (50mL), and was treated with added potassium carbonate (7.5 g) andiodomethane (8.5 mL). After stirring for 24 h the reaction mixture wasdiluted with ethyl acetate, washed with water, brine, dried (MgSO₄) andthe solvent removed in vacuo. The residue was purified by flashchromatography to yield 4-methanesulfonyl-piperazine-1,3-dicarboxylicacid 1-tert-butyl ester 3-methyl ester (3.27 g).

To a suspension of lithium aluminium hydride (0.75 g) in THF (30 mL) wasadded a solution of 4-methanesulfonyl-piperazine-1,3-dicarboxylic acid1-tert-butyl ester 3-methyl ester (3.2 g) in THF (20 mL) at 0° C. Thereaction mixture was subsequently warmed to room temperature. Afterstirring for 2.5 h, ammonium chloride solution (5 mL) was added to thereaction mixture, which was then filtered through celite, washed withbrine, dried (MgSO₄) and the solvent removed in vacuo. The residue waspurified by flash chromatography to yield3-hydroxymethyl-4-methanesulfonyl-piperazine-1-carboxylic acidtert-butyl ester (1.13 g). Treatment of this compound with HCl inDCM/MeOH yielded the desired amine, which was isolated as thehydrochloride salt.

¹H NMR (400 MHz, CDCl₃) 2.38 (1H, m), 2.50 (1H, dd, J=3.9 Hz, 7.7 Hz),2.70 (1H, br s), 2.97 (1H, m), 3.00 (3H, s), 3.06 (1H, d, J=11.6 Hz),3.52 (1H, m), 3.72 (1H, d, J=12.8 Hz), 3.86 (2H, m), 3.94 (5H, m), 4.01(1H, m), 4.10 (5H, m), 7.40 (1H, s), 7.51 (1H, t, J=7.8 Hz), 7.60 (1H,d, J=8.2 Hz); 8.27 (1H, d, J=7.2 Hz); 9.02 (1H, s); 10.16 (1H, br s). MS(ESI⁺) 544 (MH⁺).

1752-(1H-Indazol-4-yl)-6-(4-methanesulfonyl-3-methoxymethyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine

Via2-Chloro-6-(4-methanesulfonyl-3-methoxymethyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine,prepared from 1-methanesulfonyl-2-methoxymethyl-piperazine

Amine preparation: to a THF (5 mL) solution of3-hydroxymethyl-4-methanesulfonyl-piperazine-1-carboxylic acidtert-butyl ester (0.30 g, described for the production of 174) was addedsodium hydride (0.043 g). The reaction mixture was stirred for 20 minand then iodomethane (0.19 mL) was added. After stirring for 24 h thereaction mixture was then diluted with DCM, washed with brine, dried(MgSO₄) and the solvent removed in vacuo. The residue was purified byflash chromatography to yield4-methanesulfonyl-3-methoxymethyl-piperazine-1-carboxylic acidtert-butyl ester (0.254 g). Treatment of this compound with HCl inDCM/MeOH yielded the desired amine, isolated as the hydrochloride salt.

¹H NMR (400 MHz, CDCl₃): 2.34 (1H, m), 2.42 (1H, dd, J=3.9, 7.6 Hz),2.94 (2H, m), 3.07 (3H, s), 3.28 (1H, m), 3.40 (3H, s), 3.74 (1H, d,J=12.9 Hz), 3.84 (2H, m), 3.94 (4H, m), 4.00 (1H, t, J=5.1 Hz), 4.10(4H, m), 4.20 (1H, m), 7.40 (1H, s), 7.52 (1H, t, J=7.8 Hz), 7.60 (1H,d, J=8.3 Hz), 8.28 (1H, d, J=7.1 Hz), 9.00 (1H, s), 10.15 (1H, br s); MS(ESI⁺) 558 (MH⁺).

EXAMPLE 3 Biological Testing

Compounds of the invention, prepared as described in the precedingExamples, were submitted to the following series of biological assays:

(i) PI3K Biochemical Screening

Compound inhibition of PI3K was determined in a radiometric assay usingpurified, recombinant enzyme and ATP at a concentration of 1 uM. Allcompounds were serially diluted in 100% DMSO. The kinase reaction wasincubated for 1 h at room temperature, and the reaction was terminatedby the addition of PBS. IC₅₀ values were subsequently determined usingsigmoidal dose-response curve fit (variable slope). All of the compoundsexemplified had an IC₅₀ against PI3K of 510 uM or less. In particular,all of the compounds tested against the p110δ isoform of PI3K had anIC₅₀ of 0.1 μM or less.

(ii) Cellular Proliferation Inhibition

Cells were seeded at optimal density in a 96 well plate and incubatedfor 4 days in the presence of test compound. Alamar Blue™ wassubsequently added to the assay medium, and cells were incubated for 6 hbefore reading at 544 nm excitation, 590 nm emission. EC₅₀ values werecalculated using a sigmoidal dose response curve fit. All the compoundstested had an EC₅₀s of 50 uM or less in the range of cell linesutilized.

(iii) Caco-2 Permeability

Caco-2 cells were seeded onto Millipore Multiscreen plates at 1×10⁵cells/cm², and were cultured for 20 days. Assessment of compoundpermeability was subsequently conducted. The compounds were applied tothe apical surface (A) of cell monolayers and compound permeation intothe basolateral (B) compartment was measured. This was performed in thereverse direction (B-A) to investigate active transport. A permeabilitycoefficient value, P_(app), for each compound, a measure of the rate ofpermeation of the compound across the membrane, was calculated.Compounds were grouped into low (P_(app)</=1.0×10⁶ cm/s) or high(P_(app)>/=1.0×10⁶ cm/s) absorption potential based on comparison withcontrol compounds with established human absorption.

For assessment of a compound's ability to undergo active efflux, theratio of basolateral (B) to apical (A) transport compared with A to Bwas determined. Values of B−A/A−B>/=1.0 indicated the occurrence ofactive cellular efflux. All of the compounds tested through the Caco-2permeability screen had P_(app) values>/=1.0×10⁶ cm/s. One compoundassessed through the bidirectional assay, PI540, had an B−A/A−Basymmetry index of less than 1.0, indicating that the compound does notundergo active cellular efflux.

(iv) Hepatocyte Clearance

Suspensions of cryopreserved human hepatocytes were used. Incubationswere performed at compound concentration of 1 mM or 3 μM at a celldensity of 0.5×10⁶ viable cells/mL. The final DMSO concentration in theincubation was 0.25%. Control incubations were also performed in theabsence of cells to reveal any non-enzymatic degradation. Duplicatesamples (50 μL) were removed from the incubation mixture at 0, 5, 10,20, 40 and 60 minutes (control sample at 60 minutes only) and added toMeOH—containing internal standard (100 μL)—to terminate the reaction.Tolbutamide, 7-hydroxycoumarin, and testosterone were used as controlcompounds. Samples were centrifuged and the supernatants at each timepoint pooled for analysis by LC-MSMS. From a plot of ln peak area ratio(parent compound peak area/internal standard peak area) against time,intrinsic clearance (CL_(int)) was calculated as follows: CL_(int)(μl/min/million cells)=V×k, where k is the elimination rate constant,obtained from the gradient of ln concentration plotted against time; Vis a volume term derived from the incubation volume and is expressed asuL 10⁶ cells⁻¹.

Compounds were classified with low (CL</=4.6 μL/min/10⁶ cells), medium(CL>/=4.6; </=25.2 μl/min/10⁶ cells) and high (>/=25.2 μl/min/10⁶ cells)clearance. The majority of the tested compounds of the invention weredetermined to have low hepatocyte clearance.

(v) Cytochrome P450 Inhibition

Compounds of the invention were screened against five CYP450 targets(1A2, 2C9, 2C19, 2D6, 3A4) at 10 concentrations in duplicate, with a topconcentration of 100 uM being used. Standard inhibitors (furafylline,sulfaphenazole, tranylcypromine, quinidine, ketoconazole) were used ascontrols. Plates were read using a BMG LabTechnologies PolarStar influorescence mode. The majority of the tested compounds assessed in thisassay displayed weak activity (IC_(50>/=5) uM) against all isoforms ofCYP450.

(vi) Cytochrome P450 Induction

Freshly isolated human hepatocytes from a single donor were cultured for48 h prior to addition of test compound at three concentrations and wereincubated for 72 h. Probe substrates for CYP3A4 and CYP1A2 were addedfor 30 minutes and 1 h before the end of the incubation. At 72 h, cellsand media were removed and the extent of metabolism of each probesubstrate quantified by LC-MS/MS. The experiment was controlled by usinginducers of the individual P450s incubated at one concentration intriplicate. The compounds of the invention assessed in this assay showednegligible effects on induction of cytochrome P450 enzymes.

(vii) Plasma Protein Binding

Solutions of test compound (5 um, 0.5% final DMSO concentration) wereprepared in buffer and 10% plasma (v/v in buffer). A 96 well HT dialysisplate was assembled so that each well was divided in two by asemi-permeable cellulose membrane. The buffer solution was added to oneside of the membrane and the plasma solution to the other side;incubations were then conducted at 37° C. over 2 h in triplicate. Thecells were subsequently emptied, and the solutions for each batch ofcompounds were combined into two groups (plasma-free andplasma-containing) then analysed by LC-MSMS using two sets ofcalibration standards for plasma-free (6 points) and plasma-containingsolutions (7 points). The fraction unbound value for each compound wascalculated: highly protein bound compounds (>/=90% bound) had anFu</=0.1. The compounds of the invention assessed in this assay had Fuvalues>/=0.1.

(viii) hERG Channel Blockage

Compounds of the invention were evaluated for their ability to modulaterubidium efflux from HEK-294 cells stably expressing hERG potassiumchannels using established flux methodology. Cells were prepared inmedium containing RbCl and were plated into 96-well plates and grownovernight to form monolayers. The efflux experiment was initiated byaspirating the media and washing each well with 3×100 μL ofpre-incubation buffer (containing low [K⁺]) at room temperature.Following the final aspiration, 50 μL of working stock (2×) compound wasadded to each well and incubated at room temperature for 10 minutes. 50μL of stimulation buffer (containing high [K⁺]) was then added to eachwell giving the final test compound concentrations. Cell plates werethen incubated at room temperature for a further 10 minutes. 80 μL ofsupernatant from each well was then transferred to equivalent wells of a96-well plate and analysed via atomic emission spectroscopy. Compoundswere screened as 10 pt duplicate IC₅₀ curves, n=2, from a topconcentration of 1001M.

EXAMPLE 4 Tablet Composition

Tablets, each weighing 0.15 g and containing 25 mg of a compound of theinvention were manufactured as follows:

Composition for 10,000 tabletsActive compound (250 g)

Lactose (800 g)

Corn starch (415 g)Talc powder (30 g)Magnesium stearate (5 g)

The active compound, lactose and half of the corn starch were mixed. Themixture was then forced through a sieve 0.5 mm mesh size. Corn starch(10 g) is suspended in warm water (90 mL). The resulting paste was usedto granulate the powder. The granulate was dried and broken up intosmall fragments on a sieve of 1.4 mm mesh size. The remaining quantityof starch, talc and magnesium was added, carefully mixed and processedinto tablets.

EXAMPLE 5 Injectable Formulation

Formulation A Active compound 200 mg Hydrochloric Acid Solution 0.1M or4.0 to 7.0 Sodium Hydroxide Solution 0.1M q.s. to pH Sterile water q.s.to 10 mL

The compound of the invention was dissolved in most of the water (35°40° C.) and the pH adjusted to between 4.0 and 7.0 with the hydrochloricacid or the sodium hydroxide as appropriate. The batch was then made upto volume with water and filtered through a sterile micropore filterinto a sterile 10 mL amber glass vial (type 1) and sealed with sterileclosures and overseals.

Formulation B Active Compound 125 mg Sterile, Pyrogen-free, pH 7Phosphate 25 mL Buffer, q.s. to Active compound 200 mg Benzyl Alcohol0.10 g Glycofurol 75 1.45 g Water for injection q.s to 3.00 mL

The active compound was dissolved in the glycofurol. The benzyl alcoholwas then added and dissolved, and water added to 3 mL. The mixture wasthen filtered through a sterile micropore filter and sealed in sterile 3mL glass vials (type 1).

EXAMPLE 6 Syrup Formulation

Active compound 250 mg Sorbitol Solution 1.50 g Glycerol 2.00 g Sodiumbenzoate 0.005 g Flavour 0.0125 mL Purified Water q.s. to 5.00 mL

The compound of the invention was dissolved in a mixture of the glyceroland most of the purified water. An aqueous solution of the sodiumbenzoate was then added to the solution, followed by addition of thesorbital solution and finally the flavour. The volume was made up withpurified water and mixed well.

1. A compound which is a fused pyrimidine of formula (I):

wherein A represents a thiophene or furan ring; n is 1 or 2; R¹ is agroup of formula:

wherein m is 0 or 1; R³⁰ is H or C₁-C₆ alkyl; R⁴ and R⁵ form, togetherwith the N atom to which they are attached, a 5- or 6-membered saturatedN-containing heterocyclic group which includes 0 or 1 additionalheteroatoms selected from N, S and O, which may be fused to a benzenering and which is unsubstituted or substituted; or one of R⁴ and R⁵ isalkyl and the other is a 5- or 6-membered saturated N-containingheterocyclic group as defined above or an alkyl group which issubstituted by a 5- or 6-membered saturated N-containing heterocyclicgroup as defined above; R² is selected from: (a)

wherein R⁶ and R⁷ form, together with the nitrogen atom to which theyare attached, a morpholine, thiomorpholine, piperidine, piperazine,oxazepane or thiazepane group which is unsubstituted or substituted; and(b)

wherein Y is a C₂-C₄ alkylene chain which contains, between constituentcarbon atoms of the chain and/or at one or both ends of the chain, 1 or2 heteroatoms selected from O, N and S, and which is unsubstituted orsubstituted; and R³ is an indazole group which is unsubstituted orsubstituted; or a pharmaceutically acceptable salt thereof.
 2. Acompound according to claim 1 wherein the fused pyrimidine is of formula(Ia):

wherein X is S or O and R¹, R², R³ and n are as defined in claim
 1. 3. Acompound according to claim 1 wherein the fused pyrimidine is of formula(Ib):

wherein X is S or O and R¹, R², R³ and n are as defined in claim
 1. 4. Acompound according to claim 1 which is selected from:2-(1H-Indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-sulfonicacid dimethylamide;{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-morpholin-4-yl-methanone;4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-carboxylicacid (2-methoxy-ethyl)-methyl-amide;{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-N,N-dimethyl-acetamide;4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-carboxylicacid dimethylamide;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(3-morpholin-4-yl-propane-1-sulfonyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-methyl-amine;(3-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazine-1-sulfonyl}-propyl)-dimethyl-amine;2-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-2-methyl-propan-1-ol;1′-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-[1,4′]bipiperidinyl;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-morpholin-4-yl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyrimidin-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;1-(2-Hydroxy-ethyl)-4-[2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-2-one;6-(4-Cyclopropylmethyl-piperazin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-thiazol-2-yl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;2-(6-Fluoro-1H-indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-2-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-thiazol-2-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-[4-(5-methyl-furan-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylicacid amide;2-(1H-Indazol-4-yl)-6-[4-(2-methoxy-1,1-dimethyl-ethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-[(3R,5S)-4-(2-methoxy-ethyl)-3,5-dimethyl-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylicacid (2-methoxy-ethyl)-methyl-amide;1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylicacid dimethylamide;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-3-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-4-carboxylicacid methylamide;2-{4-[2-(H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-N-methyl-isobutyramide;)2-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-2-methyl-1-pyrrolidin-1-yl-propan-1-one;2-(1H-Indazol-4-yl)-6-[4-(1-methyl-1H-imidazol-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-[4-(5-methyl-isoxazol-3-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;1-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-2-methyl-propan-2-ol;Cyclopropylmethyl-{1-[2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-amine;6-[4-(1-Ethyl-1-methoxymethyl-propyl)-piperazin-1-ylmethyl]-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-[4-(1-methoxymethyl-cyclopropyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-(2,2,2-trifluoro-ethyl)-amine;2-(1H-Indazol-4-yl)-6-[4-(2-methoxy-ethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-methanol;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-4-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-[4-(6-methyl-pyridin-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-[4-(4-methyl-thiazol-2-ylmethyl)-piperazin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-pyridin-2-yl-amine;N-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-2-methoxy-N-methyl-acetamide;N-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-N-methyl-methanesulfonamide;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(3-methoxy-propyl)-methyl-amine;6-((3S,5R)-3,5-Dimethyl-4-pyridin-2-ylmethyl-piperazin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-(4-methoxymethyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(2-methoxy-ethyl)-thiazol-2-ylmethyl-amine;1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-4-pyridin-2-ylmethyl-piperidin-4-ol;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-isopropyl-(2-methoxy-ethyl)-amine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(pyridin-2-yloxy)-piperidin-1-ylmethyl]-thieno[3,2-d]pyrimidine;N-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-N-(2-methoxy-ethyl)-methanesulfonamide;2-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-propan-2-ol;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(1-oxy-pyridin-3-ylmethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-morpholin-4-ylmethyl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-ylmethyl}-(2-methoxy-ethyl)-methyl-amine;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-ylmethyl}-dimethyl-amine;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-3-yl}-(2-methoxy-ethyl)-methyl-amine;1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-3-carboxylicacid methylamide;2-(1H-Indazol-4-yl)-6-(3-methoxymethyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-pyridin-2-ylmethyl-piperidin-1-ylmethyl)-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-[4-(2-methoxy-ethoxy)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;6-((3R,5S)-3,5-Dimethyl-4-thiazol-2-ylmethyl-piperazin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-[4-(1-oxy-pyridin-2-ylmethyl)-piperazin-1-ylmethyl]-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-[4-(2-methoxy-ethyl)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-(4-methanesulfonyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-(3-methanesulfonyl-propyl)-methyl-amine;2-(1H-Indazol-4-yl)-6-[4-(3-methoxy-propane-1-sulfonyl)-piperidin-1-ylmethyl]-4-morpholin-4-yl-thieno[3,2-d]pyrimidine.(R)-1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-3-carboxylicacid methylamide;(S)-1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidine-3-carboxylicacid methylamide;6-(4-Imidazol-1-ylmethyl-piperidin-1-ylmethyl)-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-morpholin-4-ylmethyl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-6-(3-methyl-piperidin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-3-yl}-methanol;2-{1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperidin-4-yl}-ethanol;1-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-4-thiazol-2-yl-piperidin-4-ol;2-(1-Methyl-1H-indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(2-Methyl-2H-indazol-4-yl)-6-(4-methyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;152-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-(4-thiazol-4-ylmethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;1-{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-piperazin-1-yl}-3-phenoxy-propan-2-ol;6-[4-(1H-Imidazol-2-ylmethyl)-piperazin-1-ylmethyl]-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;6-[4-(3H-Imidazol-4-ylmethyl)-piperazin-1-ylmethyl]-2-(1H-indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;2-(1H-Indazol-4-yl)-4-morpholin-4-yl-6-((2S,6R)-2,4,6-trimethyl-piperazin-1-ylmethyl)-thieno[3,2-d]pyrimidine;{4-[2-(1H-Indazol-4-yl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidin-6-ylmethyl]-1-methanesulfonyl-piperazin-2-yl}-methanol;and2-(1H-Indazol-4-yl)-6-(4-methanesulfonyl-3-methoxymethyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine;and the pharmaceutically acceptable salts of the above-mentioned freecompounds.
 5. A process for producing a compound as defined in claim 1wherein m is 1, which process comprises: (a) treating a compound offormula (II):

wherein A and R² are as defined in claim 1, with a boronic acid or esterthereof of formula R³B(OR¹⁵)₂, in which R³ is as defined above and eachR¹⁵ is H or C₁-C₆ alkyl or the two groups OR¹⁵ form, together with theboron atom to which they are attached, a pinacolato boronate estergroup, in the presence of a Pd catalyst; and treating the resultingcompound of formula (III):

wherein A, R² and R³ are as defined above, with an amine of formulaNHR⁴R⁵ wherein R⁴ and R⁵ are as defined in claim 1, in the presence of asuitable reducing agent; or (b) treating a compound of formula (II) asdefined above with an amine of formula NHR⁴R⁵ wherein R⁴ and R⁵ are asdefined above, in the presence of a suitable reducing agent; andtreating the resulting compound of formula (IV):

wherein A, R², R⁴ and R⁵ are as defined above, with a boronic acid orester thereof of formula R³B(OR¹⁵)₂, in which R³ is as defined above andeach R¹⁵ is H or C₁-C₆ alkyl or the two groups OR¹⁵ form, together withthe boron atom to which they are attached, a pinacolato boronate estergroup, in the presence of a Pd catalyst.
 6. A process for producing acompound as defined in claim 1 wherein m is 0, which process comprisestreating a compound of formula (XIV):

wherein A, R² and R³ are as defined in claim 1 and W is a halo groupselected from Br and I, with an amine of formula NHR⁴R⁵ in which R⁴ andR⁵ are as defined in claim 1, in the presence of a palladium catalyst.7. A process according to claim 5 which further comprises converting theresulting compound of formula (I) into a pharmaceutically acceptablesalt thereof.
 8. A process according to claim 6 which further comprisesconverting the resulting compound of formula (I) into a pharmaceuticallyacceptable salt thereof.
 9. A pharmaceutical composition which comprisesa pharmaceutically acceptable carrier or diluent and, as an activeingredient, a compound as claimed in claim
 1. 10. A method of treating adisease or disorder arising from abnormal cell growth, function orbehaviour associated with PI3 kinase, which method comprisesadministering to a patient in need thereof a compound as defined inclaim
 1. 11. A method according to claim 10 wherein the disease ordisorder is selected from the group consisting of cancer, immunedisorders, cardiovascular disease, viral infection, inflammation,metabolism/endocrine disorders and neurological disorders.